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The patch clamp technique is a laboratory technique in electrophysiology used to study ionic currents in individual isolated living cells, tissue sections, or patches of cell membrane. The technique is especially useful in the study of excitable cells such as neurons, cardiomyocytes, muscle fibers, and pancreaticbeta cells, and can also be applied to the study of bacterial ion channels in specially prepared giant spheroplasts.
Patch clamping can be performed using the voltage clamp technique. In this case, the voltage across the cell membrane is controlled by the experimenter and the resulting currents are recorded. Alternatively, the current clamp technique can be used. In this case, the current passing across the membrane is controlled by the experimenter and the resulting changes in voltage are recorded, generally in the form of action potentials.
Erwin Neher and Bert Sakmann developed the patch clamp in the late 1970s and early 1980s. This discovery made it possible to record the currents of single ion channel molecules for the first time, which improved understanding of the involvement of channels in fundamental cell processes such as action potentials and nerve activity. Neher and Sakmann received the Nobel Prize in Physiology or Medicine in 1991 for this work.
During a patch clamp recording, a hollow glass tube known as a micropipette or patch pipette filled with an electrolyte solution and a recording electrode connected to an amplifier is brought into contact with the membrane of an isolated cell. Another electrode is placed in a bath surrounding the cell or tissue as a reference ground electrode. An electrical circuit can be formed between the recording and reference electrode with the cell of interest in between.
The solution filling the patch pipette might match the ionic composition of the bath solution, as in the case of cell-attached recording, or match the cytoplasm, for whole-cell recording. The solution in the bath solution may match the physiological extracellular solution, the cytoplasm, or be entirely non-physiological, depending on the experiment to be performed. The researcher can also change the content of the bath solution (or less commonly the pipette solution) by adding ions or drugs to study the ion channels under different conditions.
Depending on what the researcher is trying to measure, the diameter of the pipette tip used may vary, but it is usually in the micrometer range. This small size is used to enclose a cell membrane surface area or "patch" that often contains just one or a few ion channel molecules. This type of electrode is distinct from the "sharp microelectrode" used to puncture cells in traditional intracellular recordings, in that it is sealed onto the surface of the cell membrane, rather than inserted through it.
In some experiments, the micropipette tip is heated in a microforge to produce a smooth surface that assists in forming a high resistance seal with the cell membrane. To obtain this high resistance seal, the micropipette is pressed against a cell membrane and suction is applied. A portion of the cell membrane is suctioned into the pipette, creating an omega-shaped area of membrane which, if formed properly, creates a resistance in the 10–100 gigaohms range, called a "gigaohm seal" or "gigaseal". The high resistance of this seal makes it possible to isolate electronically the currents measured across the membrane patch with little competing noise, as well as providing some mechanical stability to the recording.
Many patch clamp amplifiers do not use true voltage clamp circuitry, but instead are differential amplifiers that use the bath electrode to set the zero current (ground) level. This allows a researcher to keep the voltage constant while observing changes in current. To make these recordings, the patch pipette is compared to the ground electrode. Current is then injected into the system to maintain a constant, set voltage. The current that is needed to clamp the voltage is opposite in sign and equal in magnitude to the current through the membrane.
Alternatively, the cell can be current clamped in whole-cell mode, keeping current constant while observing changes in membrane voltage.
Several variations of the basic technique can be applied, depending on what the researcher wants to study. The inside-out and outside-out techniques are called "excised patch" techniques, because the patch is excised (removed) from the main body of the cell. Cell-attached and both excised patch techniques are used to study the behavior of individual ion channels in the section of membrane attached to the electrode.
Whole-cell patch and perforated patch allow the researcher to study the electrical behavior of the entire cell, instead of single channel currents. The whole-cell patch, which enables low-resistance electrical access to the inside of a cell, has now largely replaced high-resistance microelectrode recording techniques to record currents across the entire cell membrane.
For this method, the pipette is sealed onto the cell membrane to obtain a gigaseal (a seal with electrical resistance on the order of a gigaohm), while ensuring that the cell membrane remains intact. This allows the recording of currents through single, or a few, ion channels contained in the patch of membrane captured by the pipette. By only attaching to the exterior of the cell membrane, there is very little disturbance of the cell structure. Also, by not disrupting the interior of the cell, any intracellular mechanisms normally influencing the channel will still be able to function as they would physiologically. Using this method it is also relatively easy to obtain the right configuration, and once obtained it is fairly stable.
For ligand-gated ion channels or channels that are modulated by metabotropic receptors, the neurotransmitter or drug being studied is usually included in the pipette solution, where it can interact with what used to be the external surface of the membrane. The resulting channel activity can be attributed to the drug being used, although it is usually not possible to then change the drug concentration inside the pipette. The technique is thus limited to one point in a dose response curve per patch. Therefore, the dose response is accomplished using several cells and patches. However, voltage-gated ion channels can be clamped successively at different membrane potentials in a single patch. This results in channel activation as a function of voltage, and a complete I-V (current-voltage) curve can be established in only one patch. Another potential drawback of this technique is that, just as the intracellular pathways of the cell are not disturbed, they cannot be directly modified either.
In the inside-out method, a patch of the membrane is attached to the patch pipette, detached from the rest of the cell, and the cytosolic surface of the membrane is exposed to the external media, or bath. One advantage of this method is that the experimenter has access to the intracellular surface of the membrane via the bath and can change the chemical composition of what the inside surface of the membrane is exposed to. This is useful when an experimenter wishes to manipulate the environment at the intracellular surface of single ion channels. For example, channels that are activated by intracellular ligands can then be studied through a range of ligand concentrations.
To achieve the inside-out configuration, the pipette is attached to the cell membrane as in the cell-attached mode, forming a gigaseal, and is then retracted to break off a patch of membrane from the rest of the cell. Pulling off a membrane patch often results initially in the formation of a vesicle of membrane in the pipette tip, because the ends of the patch membrane fuse together quickly after excision. The outer face of the vesicle must then be broken open to enter into inside-out mode; this may be done by briefly taking the membrane through the bath solution/air interface, by exposure to a low Ca2+ solution, or by momentarily making contact with a droplet of paraffin or a piece of cured silicone polymer.
Whole-cell recording or whole-cell patch
Whole-cell recordings involve recording currents through multiple channels simultaneously, over a large region of the cell membrane. The electrode is left in place on the cell, as in cell-attached recordings, but more suction is applied to rupture the membrane patch, thus providing access from the interior of the pipette to the intracellular space of the cell. This provides a means to administer and study how treatments (e.g. drugs) can affect cells in real time. Once the pipette is attached to the cell membrane, there are two methods of breaking the patch. The first is by applying more suction. The amount and duration of this suction depends on the type of cell and size of the pipette. The other method requires a large current pulse to be sent through the pipette. How much current is applied and the duration of the pulse also depend on the type of cell. For some types of cells, it is convenient to apply both methods simultaneously to break the patch.
The advantage of whole-cell patch clamp recording over sharp electrode technique recording is that the larger opening at the tip of the patch clamp electrode provides lower resistance and thus better electrical access to the inside of the cell. A disadvantage of this technique is that because the volume of the electrode is larger than the volume of the cell, the soluble contents of the cell's interior will slowly be replaced by the contents of the electrode. This is referred to as the electrode "dialyzing" the cell's contents. After a while, any properties of the cell that depend on soluble intracellular contents will be altered. The pipette solution used usually approximates the high-potassium environment of the interior of the cell to minimize any changes this may cause. There is often a period at the beginning of a whole-cell recording when one can take measurements before the cell has been dialyzed.
The name "outside-out" emphasizes both this technique's complementarity to the inside-out technique, and the fact that it places the external rather than intracellular surface of the cell membrane on the outside of the patch of membrane, in relation to the patch electrode.
The formation of an outside-out patch begins with a whole-cell recording configuration. After the whole-cell configuration is formed, the electrode is slowly withdrawn from the cell, allowing a bulb of membrane to bleb out from the cell. When the electrode is pulled far enough away, this bleb will detach from the cell and reform as a convex membrane on the end of the electrode (like a ball open at the electrode tip), with the original outside of the membrane facing outward from the electrode. As the image at the right shows, this means that the fluid inside the pipette will be simulating the intracellular fluid, while a researcher is free to move the pipette and the bleb with its channels to another bath of solution. While multiple channels can exist in a bleb of membrane, single channel recordings are also possible in this conformation if the bleb of detached membrane is small and only contains one channel.
Outside-out patching gives the experimenter the opportunity to examine the properties of an ion channel when it is isolated from the cell and exposed successively to different solutions on the extracellular surface of the membrane. The experimenter can perfuse the same patch with a variety of solutions in a relatively short amount of time, and if the channel is activated by a neurotransmitter or drug from the extracellular face, a dose-response curve can then be obtained. This ability to measure current through exactly the same piece of membrane in different solutions is the distinct advantage of the outside-out patch relative to the cell-attached method. On the other hand, it is more difficult to accomplish. The longer formation process involves more steps that could fail and results in a lower frequency of usable patches.
This variation of the patch clamp method is very similar to the whole-cell configuration. The main difference lies in the fact that when the experimenter forms the gigaohm seal, suction is not used to rupture the patch membrane. Instead, the electrode solution contains small amounts of an antifungal or antibiotic agent, such as amphothericin-B, nystatin, or gramicidin, which diffuses into the membrane patch and forms small pores in the membrane, providing electrical access to the cell interior. When comparing the whole-cell and perforated patch methods, one can think of the whole-cell patch as an open door, in which there is complete exchange between molecules in the pipette solution and the cytoplasm. The perforated patch can be likened to a screen door that only allows the exchange of certain molecules from the pipette solution to the cytoplasm of the cell.
Advantages of the perforated patch method, relative to whole-cell recordings, include the properties of the antibiotic pores, that allow equilibration only of small monovalent ions between the patch pipette and the cytosol, but not of larger molecules that cannot permeate through the pores. This property maintains endogenous levels of divalent ions such as Ca2+ and signaling molecules such as cAMP. Consequently, one can have recordings of the entire cell, as in whole-cell patch clamping, while retaining most intracellular signaling mechanisms, as in cell-attached recordings. As a result, there is reduced current rundown, and stable perforated patch recordings can last longer than one hour. Disadvantages include a higher access resistance, relative to whole-cell, due to the partial membrane occupying the tip of the electrode. This may decrease current resolution and increase recording noise. It can also take a significant amount of time for the antibiotic to perforate the membrane (about 15 minutes for amphothericin-B, and even longer for gramicidin and nystatin). The membrane under the electrode tip is weakened by the perforations formed by the antibiotic and can rupture. If the patch ruptures, the recording is then in whole-cell mode, with antibiotic contaminating the inside of the cell.
a loose patch clamp is different from the other techniques discussed here in that it employs a loose seal (low electrical resistance) rather than the tight gigaseal used in the conventional technique. This technique was used as early as the year 1961, as described in a paper by Strickholm on the impedance of a muscle cell's surface, but received little attention until being brought up again and given a name by Almers, Stanfield, and Stühmer in 1982, after patch clamp had been established as a major tool of electrophysiology.
To achieve a loose patch clamp on a cell membrane, the pipette is moved slowly towards the cell, until the electrical resistance of the contact between the cell and the pipette increases to a few times greater resistance than that of the electrode alone. The closer the pipette gets to the membrane, the greater the resistance of the pipette tip becomes, but if too close a seal is formed, and it could become difficult to remove the pipette without damaging the cell. For the loose patch technique, the pipette does not get close enough to the membrane to form a gigaseal or a permanent connection, nor to pierce the cell membrane. The cell membrane stays intact, and the lack of a tight seal creates a small gap through which ions can pass outside the cell without entering the pipette.
A significant advantage of the loose seal is that the pipette that is used can be repeatedly removed from the membrane after recording, and the membrane will remain intact. This allows repeated measurements in a variety of locations on the same cell without destroying the integrity of the membrane. This flexibility has been especially useful to researchers for studying muscle cells as they contract under real physiological conditions, obtaining recordings quickly, and doing so without resorting to drastic measures to stop the muscle fibers from contracting. A major disadvantage is that the resistance between the pipette and the membrane is greatly reduced, allowing current to leak through the seal, and significantly reducing the resolution of small currents. This leakage can be partially corrected for, however, which offers the opportunity to compare and contrast recordings made from different areas on the cell of interest. Given this, it has been estimated that the loose patch technique can resolve currents smaller than 1 mA/cm2.
Automatic patch clamping
Automated patch clamp systems have been developed in order to collect large amounts of data inexpensively in a shorter period of time. Such systems typically include a single-use microfluidic device, either an injection molded or a polydimethylsiloxane (PDMS) cast chip, to capture a cell or cells, and an integrated electrode.
In one form of such an automated system, a pressure differential is used to force the cells being studied to be drawn towards the pipette opening until they form a gigaseal. Then, by briefly exposing the pipette tip to the atmosphere, the portion of the membrane protruding from the pipette bursts, and the membrane is now in the inside-out conformation, at the tip of the pipette. In a completely automated system, the pipette and the membrane patch can then be rapidly moved through a series of different test solutions, allowing different test compounds to be applied to the intracellular side of the membrane during recording.
- ^"The Nobel Prize in Physiology or Medicine 1991". nobelprize.org. Nobel Media AB. Retrieved November 8, 2014.
- ^Bannister, Niel (November 1, 2012). Langton, Phil (ed.). Essential Guide to Reading Biomedical Papers: Recognizing and Interpreting Best Practice. Wiley-Blackwell. doi:10.1002/9781118402184. ISBN .
- ^ abcdSakmann, B.; Neher, E. (1984). "Patch clamp techniques for studying ionic channels in excitable membranes". Annual Review of Physiology. 46: 455–472. doi:10.1146/annurev.ph.46.030184.002323. hdl:21.11116/0000-0000-D552-3. PMID 6143532.
- ^Sigworth, Fredrick J.; Neher, E. (October 2, 1980). "Single Na+ channel currents observed in cultured rat muscle cells". Nature. 287 (5781): 447–449. Bibcode:1980Natur.287..447S. doi:10.1038/287447a0. PMID 6253802. S2CID 4238010.
- ^Ellen Covey; Matt Carter (2015). Basic Electrophysiological Methods. Oxford University Press. pp. 22–. ISBN .
- ^ abcdeMolleman, Areles (March 6, 2003). Patch Clamping: An Introductory Guide To Patch Clamp Electrophysiology. Wiley. doi:10.1002/0470856521. ISBN .
- ^Veitinger, Sophie (2011-11-09). "The Patch-Clamp Technique". Science Lab. Leica Microsystems. Retrieved November 10, 2014.
- ^Ogden, David; Stanfield, Peter. "Patch Clamp Techniques"(PDF). utdallas.edu. pp. 53–78. Retrieved November 11, 2014.
- ^ abSegev, Amir; Garcia-Oscos, Francisco; Kourrich, Saïd (2016-06-15). "Whole-cell Patch-clamp Recordings in Brain Slices". Journal of Visualized Experiments (112): e54024. doi:10.3791/54024. ISSN 1940-087X. PMC 4927800. PMID 27341060.
- ^Staley, K.J.; Otis, T. S.; Mody, I (May 1, 1992). "Membrane properties of dentate gyrus granule cells: comparison of sharp microelectrode and whole-cell recordings". Journal of Neurophysiology. 67 (5): 1346–1358. doi:10.1152/jn.19188.8.131.526. PMID 1597717.
- ^Howe, JR; Cull-Candy, SG; Colquhoun, D (Jan 1991). "Currents through single glutamate receptor channels in outside-out patches from rat cerebellar granule cells". Journal of Physiology. 432 (1): 143–202. doi:10.1113/jphysiol.1991.sp018381. PMC 1181322. PMID 1715916.
- ^ abcLinley, John (2013). "Perforated Whole-Cell Patch-Clamp Recording". In Gamper, Nikita (ed.). Ion Channels. Methods in Molecular Biology. 998 (Second ed.). Humana Press. pp. 149–157. doi:10.1007/978-1-62703-351-0_11. ISBN . PMID 23529427.
- ^Strickholm, A (1 Jul 1961). "Impedance of a Small Electrically Isolated Area of the Muscle Cell Surface". Journal of General Physiology. 44 (6): 1073–88. doi:10.1085/jgp.44.6.1073. PMC 2195146. PMID 19873540.
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- ^ abLupa, MT; Caldwell, JH (Nov 1991). "Effect of Agrin on the Distribution of Acetylcholine Receptors and Sodium Channels on Adult Skeletal Muscle Fibers in Culture". Journal of Cell Biology. 115 (3): 765–778. doi:10.1083/jcb.115.3.765. PMC 2289169. PMID 1655812.
- ^Bowlby, Mark; Merrill, Thomas; Vasilyev, Dmitry (2005). "Development of a Novel Automated Ion Channel Recording Method Using Inside-Out Whole-Cell Membranes". Journal of Biomolecular Screening. 10 (8): 806–813. doi:10.1177/1087057105279481. PMID 16234349.
Intel® Management Engine Critical Firmware Update (Intel-SA-00086)
Intel® Management Engine (Intel® ME 6.x/7.x/8.x/9.x/10.x/11.x), Intel® Trusted Execution Engine (Intel® TXE 3.0), and Intel® Server Platform Services (Intel® SPS 4.0) vulnerability (Intel-SA-00086)
|Note||This article describes issues related to security vulnerabilities found in the Intel® Management Engine Firmware. This article doesn't contain information related to the processor side-channel vulnerability (known as Meltdown/Spectre). If you're looking for information on the Meltdown/Spectre issue, go to Side-Channel Analysis Facts and Intel® Products.|
In response to issues identified by external researchers, Intel has performed an in-depth comprehensive security review of the following with the objective of enhancing firmware resilience:
- Intel® Management Engine (Intel® ME)
- Intel® Trusted Execution Engine (Intel® TXE)
- Intel® Server Platform Services (SPS)
Intel has identified security vulnerabilities that could potentially impact certain PCs, servers, and IoT platforms.
Systems using Intel ME Firmware versions 6.x-11.x, servers using SPS Firmware version 4.0, and systems using TXE version 3.0 are impacted. You may find these firmware versions on certain processors from the:
- 1st, 2nd, 3rd, 4th, 5th, 6th, 7th, and 8th generation Intel® Core™ Processor Families
- Intel® Xeon® Processor E3-1200 v5 and v6 Product Family
- Intel® Xeon® Processor Scalable Family
- Intel® Xeon® W Processor
- Intel Atom® C3000 Processor Family
- Apollo Lake Intel Atom® Processor E3900 series
- Apollo Lake Intel® Pentium® Processors
- Intel® Pentium® Processor G Series
- Intel® Celeron® G, N, and J series Processors
To determine if the identified vulnerabilities impact your system, download and run the Intel CSME Version Detection tool using the links below.
Frequently Asked Questions Section
Resources for Microsoft and Linux* users
Resources from system/motherboard manufacturers
|Note||Links for other system/motherboard manufacturers will be provided when available. If your manufacturer is not listed, contact them for information on the availability of the necessary software update.|
Frequently asked questions:
Q: The Intel CSME Version Detection Tool reports that my system is vulnerable. What do I do?
A: Intel has provided system and motherboard manufacturers with the necessary firmware and software updates to resolve the vulnerabilities identified in Security Advisory Intel-SA-00086.
Contact your system or motherboard manufacturer regarding their plans for making the updates available to end users.
Some manufacturers have provided Intel with a direct link for their customers to obtain additional information and available software updates (Refer to the list below).
Q: Why do I need to contact my system or motherboard manufacturer? Why can’t Intel provide the necessary update for my system?
A: Intel is unable to provide a generic update due to management engine firmware customizations performed by system and motherboard manufacturers.
Q: My system is reported as may be Vulnerable by the Intel CSME Version Detection Tool. What do I do?
A: A status of may be Vulnerable is usually seen when either of the following drivers aren't installed:
- Intel® Management Engine Interface (Intel® MEI) driver
- Intel® Trusted Execution Engine Interface (Intel® TXEI) driver
Contact your system or motherboard manufacturer to obtain the correct drivers for your system.
Q: My system or motherboard manufacturer is not shown in your list. What do I do?
A: The list below shows links from system or motherboard manufacturers who have provided Intel with information. If your manufacturer is not shown, contact them using their standard support mechanisms (website, phone, email, and so on) for assistance.
Q: What types of access would an attacker need to exploit the identified vulnerabilities?
A: If the equipment manufacturer enables Intel-recommended Flash Descriptor write protections, an attacker needs physical access to platform’s firmware flash to exploit vulnerabilities identified in:
End of manufacturing
The attacker gains physical access by manually updating the platform with a malicious firmware image through a flash programmer physically connected to the platform’s flash memory. Flash Descriptor write-protection is a platform setting usually set at the end of manufacturing. Flash Descriptor write-protection protects settings on the Flash from being maliciously or unintentionally changed after manufacturing is completed.
If the equipment manufacturer doesn't enable Intel-recommended Flash Descriptor write protections, an attacker needs Operating kernel access (logical access, Operating System Ring 0). The attacker needs this access to exploit the identified vulnerabilities by applying a malicious firmware image to the platform through a malicious platform driver.
The vulnerability identified in CVE-2017-5712 is exploitable remotely over the network in conjunction with a valid administrative Intel® Management Engine credential. The vulnerability is not exploitable if a valid administrative credential is unavailable.
If you need further assistance, contact Intel Customer Support to submit an online service request.
Here’s how to fix laptop battery drain after shutdown
Windows & Software Expert
Milan has been enthusiastic about PCs ever since his childhood days, and this led him to take interest in all PC-related technologies. Before joining WindowsReport, he worked as a front-end web developer. Read more
- If your laptop battery is draining while the device isn't powered on, you need a rapid fix.
- With one exception, the solutions below will only take a couple of minutes so try them out.
- In our Laptop & PC section, you'll find step-by-step guidelines for any error you might face.
- We also offer precious advice on the best tools to optimize your devices in the Optimization Hub.
Powering off your laptop should end any running process on it. Yet, users noticed the battery drain even after shutdown.
For instance, many complained that their laptop battery drains to 0% while powered off. This is a fairly common problem, and it’s usually caused by the Connected Standby feature.
Then, some say that laptop battery self-discharges. This is a cause of an improper shutdown and can be fixed by pressing and holding the Power button to turn off the device completely.
The issue affects any laptop brand, however, it’s been reported more often for Surface Pro 4 and HP.
To help you with this problem, we’ve compiled a list of solutions so make sure to try them one by one.
What can I do if my laptop battery is draining?
- Disable the Connected Standby feature
- Check your battery health
- Press and hold the power button for 15 seconds
- Install an older Intell driver
- Use Command Prompt to shut down your PC
- Perform a BIOS update
- Disable Fast Startup
1. Disable the Connected Standby feature
Note: This solution involves modifying the registry. Since this can be potentially dangerous, it’s advised to create a System Restore point just in case.
- Press Windows Key + R and enter regedit. Now press Enter or click OK.
- In the left pane go to the following key:
- In the right pane, double-click the CsEnabled DWORD.
- Set the Value Data to 0 and click OK to save changes.
After doing that, restart your PC and the problem should be permanently resolved. Bear in mind that by disabling this feature you’ll disable the Sleep feature completely, but at least the problem will be resolved.
Many laptops use the Connected Standby feature, and this feature won’t completely turn off your laptop. Instead, it will keep it in a state similar to Sleep Mode and allow you to easily boot your PC and continue where you left off.
Although this feature can be useful, it will drain your battery even while your laptop is off.
Interested in extending your laptop battery life? Here are 13 tips to do that.
2. Check your battery health
Another problem might be the battery health.
As you might know, each battery has a limited number of power cycles. This means that the battery can be charged and drained only a number of times before it starts losing its charge and capacity.
So if your battery has reached its charging limit cycle, it’s possible that issues start occurring.
To fix this problem, it’s advised that you replace your laptop battery and check if that helps. Before that, check your laptop battery health first.
If the battery is nearing its power cycle limit, perhaps this is a good time to replace it.
3. Press and hold the power button for 15 seconds
Many users reported that their battery drains almost halfway in a span of a few hours, even if their device is shut down.
To fix this problem, users are suggesting to press and hold the Power button for about 15 seconds after your device shuts down. This will force your laptop to go into deep shutdown mode. After doing that, your laptop will be completely turned off and there won’t be any battery drain issues.
Bear in mind that this is just a workaround, so you’ll have to repeat it every time you shut down your device.
4. Install an older Intel driver
According to users, sometimes battery drain issues can be caused by your driver, most notably by Intel Management Interface. Therefore, it’s advised that you remove it and install the older version.
On the contrary, the battery issue can also appear if some drivers are out of date. To fix that, update all major drivers on your PC.
If you know what you’re looking for, visit your laptop manufacturer’s website and download the appropriate driver.
Since this can be a tedious process, especially if you need to update several drivers, it might be useful to use a third-party tool such as DriverFix, that is set to download and install the suitable Intel driver.
This tool can particularly help with choosing the compatible software for your device, but also with pointing out which of your system components need an update.
DriverFix searches a large database of the most recent drivers and provides a range of trustworthy drivers from which you can choose, safely and fast.
The update process itself simply takes a few minutes, and the end result is fairly reassuring, confirming that you have the right driver.
5. Use Command Prompt to shut down your PC
- Type Command Prompt in the search bar and open it.
- Now run shutdown /s command and your PC will shut down completely.
Another faster method is to use the Run dialog. To do that, follow these steps:
- Press Windows Key + R to open the Run dialog.
- Enter shutdown /s command and press Enter or click OK.
If these methods work, you can speed up this process by creating a desktop shortcut and use it to shut down your PC. To do that, follow these steps:
- Right-click the Desktop and choose New > Shortcut.
- Enter shutdown /s in the input field and click Next.
- Enter the name of the new shortcut and follow the instructions on the screen.
Once you create a new shortcut, use it to turn off your PC. Again, this is just a workaround, so you’ll have to use it every ispring crack key - Crack Key For U you want to turn off your PC.
6. Perform a BIOS update
BIOS is an integral part of any PC and if your battery drains while the PC is shut down, the problem might lie here.
Several users managed to fix the battery problem simply by updating BIOS to the latest version or by reinstalling their current BIOS version.
This is an advanced procedure, and we wrote a simple guide on how to flash your BIOS. This is just a general guide, but if you want detailed instructions on how to upgrade your BIOS be sure to check your laptop manual for detailed instructions.
Once the BIOS is up to date, check if the problem is still there.
7. Disable Fast Startup
- Press Windows Key + S and enter power settings.
- Choose Power & sleep settings from the search results.
- In the right pane, scroll down to the Related settings section and click Additional power settings.
- In the left pane, select Choose what the power button does.
- Click on Change settings that are currently unavailable.
- Disable Turn on fast startup (recommended) option and click Save changes button.
After doing that, the Fast Startup feature should be disabled and your problem should be resolved.
By disabling this feature your PC might boot slower, but at least the battery drain will be resolved.
We hope that at least one of the solutions presented above worked for you. If you managed to troubleshoot the problem using another method, share it with us in the comments below.
Example: Device# no debug license errors
Disables license debugging activity on a device.
Configuring Examples for Software Licensing
Example: Installing and Upgrading Licenses
The following example shows how to use the license install command to install a license saved in TFTP on the device. The display is truncated for easier readability:Device# Installing licenses from "tftp://infra-sun/<user>/license/5400/38a.lic" Loading <user>/license/5400/38a.lic from 172.19.211.47 (via GigabitEthernet0/0): ! [OK - 1192 bytes] Extension licenses are being installed in the device with UDI "AS54XM-AC-RPS:JAE0948QXKD" for the following features: Feature Name: gsmamrnb-codec-pack PLEASE READ THE FOLLOWING TERMS CAREFULLY. . ACCEPT? [yes/no]: Issue 'license feature gsmamrnb-codec-pack' command to enable the license Installing.Feature:gsmamrnb-codec-pack.Successful:Supported
Example: Adding a Comment to a License File
The following example shows how to use the license comment command to add or delete information about a specific license. The command checks that a license associated with the specified feature is present in license storage. If a switch number is specified, this command is executed on the specified switch.
As the example shows, when the license is present and multiple license lines are stored, you are prompted to select a license line. This action helps to distinguish licenses. Type the number at the Select Index to Add Comment prompt to select the license.Device# Feature: gsmamrnb-codec-pack 1 License Type: Evaluation License State: Inactive Evaluation total period: 20 hours 0 minute Evaluation period left: 20 hours 0 minute License Addition: Additive Comment: Store Index: 0 Store Name: Primary License Storage 2 License Type: Permanent License State: Active, Not in Use License Addition: Exclusive Comment: Store Index: 1 Store Name: Primary License Storage Select Index to Add Comment [1-2]: % Success: Adding comment "Use this permanent license" succeeded Device# License Store: Primary License Storage Store Index: 0 License: 11 gsmamrnb-codec-pack 1.0 LONG TRIAL DISABLED 20 DISABLED STANDA LONE ADD INFINITE_KEYS INFINITE_KEYS NEVER NEVER NiL SLM_CODE CL_ ND_LCK NiL *1YCHJRBMWKZAED2400 NiL NiL NiL 5_MINS <UDI><PID>AS54X M-AC-RPS</PID><SN>JAE0948QXKD</SN></UDI> ,Jx8qaVf:iXWaH9PsXjkVnmz 7gWh:cxdf9nUkzY6o8fRuQbu,7wTUz237Cz6g9VjfrCk,0a2Pdo,Ow6LWxcCRFL:x cTxwnffn9i,4,aUWv8rL50opDUdAsFnxLsvoFRkcAfm$<WLC>AQEBIQAB//9NA+1m Uwfs/lD0dmdF9kyX8wDrua1TZhnnAy6Mxs1dTboIcRaahKxJJdj4Oi1w3wscqvPiA mWSaEmUT56rstk6gvmj+EQKRfD9A0ime1czrdKxfILT0LaXT416nwmfp92Tya6vIQ crack application website - Activators Patch 4FnlBdqJ1sMzXeSq8PmVcTU9A4o9hil9vKur8N9F885D9GVF0bJHciT5M=</WLC> Comment: Use this permanent license. Hash: E1WjIQo4qsl9g8cpnpoogP/0DeY= Note Book - Activators Patch
Example: Saving All Licenses to a Specified Storage Area
The following example shows how to use the license save command to save copies of all licenses to the flash file system:Device# license lines saved . to flash:all_licenses.lic
Example: Removing Licenses
The following examples Note Book - Activators Patch how to use the license clear command to remove a license entry from license storage once it has been verified that the license line is valid and was explicitly installed.
You must select the index number of the license to clear. Type the number at the Select Index to Clear prompt as shown in this example.Device# Feature: standard 1 License Type: Evaluation License State: Inactive Evaluation total period: 20 hours 0 minute Evaluation period left: 20 hours 0 minute License Addition: Additive Comment: Store Index: 0 Store Name: Primary License Storage 2 License Type: Permanent License State: Active, Not in Use License Addition: Exclusive Comment: Store Index: 1 Store Name: Primary License Storage Select Index to Clear [1-2]: Are you sure you want to clear? (yes/[no]): Device# Feature: premium Note Book - Activators Patch Period left: 1 hour 0 minute Index: 1 Feature: premium Version: 1.0 License Type: Evaluation License State: Active, Not in Use, EULA not accepted Evaluation total period: 1 hour 0 minute Evaluation period left: 1 hour 0 minute License Count: Non-Counted License Priority: None Store Index: 0 Store Name: Evaluation License Storage
Example: Rehosting (Revoking and Transferring) a License
The following example shows how to use the license revoke command to revoke a license stored in TFTP and how to transfer it to a license stored in flash memory. You might need to read and accept the terms and conditions of the license type being transferred. The following example is truncated for readability:Device# Following Permanent license(s) will be revoked from this device Feature Name: gsmamrnb-codec-pack Following Extension license(s) will be installed in this device Feature Name: gsmamrnb-codec-pack PLEASE READ THE FOLLOWING TERMS CAREFULLY. . ACCEPT? [yes/no]: Issue 'license feature gsmamrnb-codec-pack' command to enable the license Rehost ticket saved . to flash:rt.lic
Example: Generic Command Enhanced with Licensing Information
The generic commands described in the following sections are enhanced with licensing information:
The reload command shows the expired licenses, followed by expiring Note Book - Activators Patch sorted by the period left and end date:Device# The following license(s) are expiring or have expired. Features with expired licenses may not work after Reload. Feature: uc,Status: expiring, Period Left: 7 wks 5 days Proceed with reload? [confirm]
The show running-config command displays the unique device identifier (UDI) of a device. If the configuration file was copied from a different device, a warning is displayed upon reload. A UDI mismatch warning is also displayed during reload if the startup-config file has a different UDI than the platform UDI.Device# Building configuration. Current configuration : 4772 bytes ! version 12.4 no service pad service timestamps debug datetime msec service timestamps log datetime msec no service password-encryption service internal ! hostname csl-xfr-enhance-2951 ! . . license udi pid CISCO2951 sn FHH1211P037 license boot module c2951 technology-package securityk9 disable license boot module c2951 technology-package uc license boot module c2951 technology-package data license call-home url https://tools-stage.cisco.com/SWIFT/Licensing license agent listener http plaintext /lic-agent authenticate none ! ! archive log config hidekeys !. . .
The show tech-support command displays the output of the show license udi, show license file, show license detail, show license status, and the show license statistics commands.Device# 0 LONG TRIAL DISABLED 1440 DISABLED STANDALONE AD D INFINITE_KEYS INFINITE_KEYS NEVER NEVER NiL SLM_CODE DEMO NiL N iL Ni NiL NiL 5_MINS NiL GT5YVbrMAdt0NY50UcKGfvLTjQ17P2o3g84hE8Tq sOfu3Xph0N:2AmMdpMNxxKXSVG$<WLC>AQEBIQAB//+FugzZgqFJn/XhIxoyelg63 YJD++i6Qx6vVp0MVqrX2EinbufbTfGzc7/GHNZaDZqRqwInXo3s+nsLU7rOtdOxoI xYZAo3LYmUJ+MFzsqlhKoJVlPyEvQ8H21MNUjVbhoN0gyIWsyiJaM8AQIkVBQFzhr 10GYolVzdzfJfEPQIx6tZ++/Vtc/q3SF/5Ko8XCY=</WLC> Comment: Hash: CLWUVZgY84BMRTO3JIlYmIqwAQA= ------------------ show license detail ------------------ Index: 1 Feature: SNASw ant download manager addon - Crack Key For U Version: 1. ------------------ show license statistics ------------------ Administrative statistics Install success count: 0 Install failure count: 0 Install duplicate count: 0 Comment add count: 0 Comment delete count: 0 Clear count: 0 Save count: 0 Note Book - Activators Patch Save cred count: 1 Client statistics Request success count: 1 Request failure count: 3 Release count: 0 Global Notify count: 4
The show version command displays the license UDI information:Device> Cisco IOS Software, C2951 Software (C2951-UNIVERSALK9-M), Experimental Version 12.4(20090326:052343) [rifu-xformers_3_25 130] Copyright (c) 1986-2009 by Cisco Systems, Inc. Compiled Thu 26-Mar-09 21:49 by rifu ROM: System Bootstrap, Version 12.4(20090303:092436) [BLD-xformers_dev.XFR_20090303-20090303_0101-53 107], DEVELOPMENT SOFTWARE csl-xfr-enhance-2951 uptime is 3 days, 4 hours, 28 minutes System returned to ROM by reload at 18:48:45 PST Mon Nov 26 1956 System image file is "flash0:c2951-universalk9-mz.SSA" Last reload reason: Reload Command . . Cisco C2951 (revision 1.0) with 1005568K/43008K bytes of memory. Processor board ID FHH1211P037 3 Gigabit Ethernet interfaces 1 terminal line 1 cisco Special Services Engine(s) DRAM configuration is 72 bits wide with parity enabled. 255K bytes of non-volatile configuration memory. 250880K bytes of ATA System CompactFlash 0 (Read/Write) License Info: License UDI: ------------------------------------------------- Device# PID SN ------------------------------------------------- *0 CISCO2951 Note Book - Activators Patch FHH1211P037 Technology Package License Information for Module:'c2951' ---------------------------------------------------------------- Technology Technology-package Technology-package Current Type Next reboot ----------------------------------------------------------------- ipbase ipbasek9 None ipbasek9 security disable None disable uc uc Evaluation uc data None None None Configuration register is 0x0
Cisco License Manager application
User Guide for Cisco License Manager
Software activation conceptual overview
“Cisco IOS Software Activation Conceptual Overview” module
Software activation commands
Software Activation Command Reference
Cisco IOS commands
Master Commands List, All Releases
Integrated Services Routers licensing
Software Activation on Cisco Integrated Services Routers
To locate and download MIBs for selected platforms, Cisco software releases, and feature sets, use Cisco MIB Locator found at the following URL:
The Cisco Support and Documentation website provides online resources to download documentation, software, and tools. Use these resources to install and configure the software and to troubleshoot and resolve technical issues with Cisco products and technologies. Access to most tools on the Cisco Support and Documentation website requires a Cisco.com user ID and password.
Feature Information for Cisco IOS Software Activation
The following table provides release information about the feature or features described in this module. This table lists only the software release that introduced support for a given feature in a given software release train. Unless noted otherwise, subsequent releases of that software release train also support that feature.Use Cisco Feature Navigator to find information about platform support and Cisco software image support. To access Cisco Feature Navigator, go to . An account on Cisco.com is not required.
Cisco IOS Software Activation
Cisco IOS Software Activation EXEC commands support basic licensing processes.
This feature is platform-independent.
These commands were introduced or modified by this feature: debug license, license clear, license comment, license install, license revoke, license save, license save credential, show license all, show license detail, show license feature, show license file, show license statistics, show license status, show license udi
In Cisco IOS Release 15.4(1)S, support was added for the Cisco ASR 901S series router.
CISL-SNMP support (MIB)
SNMP support for the CISCO-LICENSE-MGMT-MIB was added.
These commands were introduced or modified by this feature: snmp-server enable traps, snmp-server host