You (either: "[[fill your glass electrode|fill1]]", "[[fill your glass electrode|fill3]]", "[[fill your glass electrode|fill2]]") with internal solution.(either: "The filter on your syringe seems to have become clogged. You press the plunger harder, trying to press the solution through.", "You press down on the syringe plunger but nothing happens. You try harder.", "Something's clogged. Nothing's coming out of the pipette tip. You press harder.")
(either: "The filter pops off the syringe, sending an explosion of solution into your face. You hope there's nothing too dangerous in it.", "Still nothing happens. You press the plunger against the desk. All of the solution in the syringe spurts out.")
(either: "Bloody hell", "Shit", "Darn", "Jesus Christ")!
(either: "You thaw another aliquot of internal solution", "You prepare another syringe", "You clean up") and [[try again.|start]](either: "You push the tip of the pipette a bit too far down the electrode and end up breaking the electrode.", "You clumsily try to insert the pipette into the electrode and keep missing. Your hand shakes and you accidentally break the tip of the electrode.", "You fumble and drop the electrode.", "You notice that the electrode tip is improperly formed.")
(either: "God damn it", "Whoops", "Shit", "Argh")!
(either: "You pick up a", "You get a", "Time for a") [[new electrode.|start]]You insert the pipette tip into the electrode and depress the syrnge plunger to deposit a bit of internal solution.
(either: "Great!", "That wasn't so hard!", "You've done it a million times.")
Now you can put it on (either: "[[the electrode holder|headst1]]","[[the electrode holder|headst2]]").(either: "Alas, your delicate glass electrode is several centimetres away from the headstage when you accidentally touch the tip on the microscope stage.", "On its way to the electrode holder, the tip brushes against the microscope stage and breaks.", "Crap, you just accidentally touched and broke the electrode tip.", "You place the electrode in the holder and apply positive pressure. Solution drips out of the tip. It seems the electrode was broken.")
(either: "Life is hard.", "What did you do to deserve this?", "Seriously?")
(either: "You'll have to", "Time to", "All you can do is") fill a [[new electrode.|start]](either: "You insert the electrode wire into your glass pipette and push it up into the electrode holder.", "You place the electrode in the holder and tighten the washer.", "You put the electrode on the electrode holder.")
Everything seems (either: "secure", "good", "ready").
You (either: "apply positive pressure", "blow into the mouthpiece") and (either: "[[lower the electrode into the bath.|bath1]]", "[[lower the electrode into the bath.|bath2]]")(either: "Your electrode is safely in the bath and you've located it with the microscope.", "You touch the tip of your electrode to the surface of your bath solution. You look down the microscope and easily locate it.", "The electrode is in the bath and in the middle of the microscope's field of view.")
Now it's time to (either: "[[pick a cell|cell1]]", "[[pick a cell|cell2]]", "[[pick a cell|cell3]]").(either: "You are a bit too quick when lowering the electrode, and you smash it against the bottom of the bath.", "You enthusiastically lower the electrode into the bath. You forget to change the manipulator speed and send the electrode crashing into the bottom of the bath.", "The electrode flies into the bottom of the bath and breaks.")
(either: "Whoops", "Shit", "You hate this microscope", "Stupid manipulator").
(either: "Time to prepare", "You get",) a [[new electrode.|start]]Well, it's (either: "not the happiest looking cell", "a bit crunchy-looking", "kind of small", "weirdly shaped"), but (either: "it's slim pickings on this coverslip", "you're desperate for data", "you know it wasn't the greatest prep"). You shrug and lower your electrode onto it.
When you see the tip resistance increase slightly, you release the positive pressure and (either: "[[mouth pipette it up|seal1]]", "[[mouth pipette|seal2]]", "[[mouth pipette|seal3]]").(either: "There's a nice looking cell.", "There's a nice big cell.", "You decide to just pick the cell that's closest to your electrode.") You slowly move your electrode tip towards it.
(either: "As your electrode approaches it, it starts to disintegrate.", "You see a dimple on the surface and move to release the pressure, but you're too slow and the cell explodes.") (either: "Damn!", "Shoot.", "Crud.")
Time to [[start again|start]].Look at that (either: "beautiful", "perfect", "gorgeous") cell. (either: "You almost don't want to ruin it by lowering your electrode onto it", "You're going to kill it", "It looks like it'll give you data").
When (either: "you see", "you think you can see", "you sort of see") a dimple form on its surface, you release your positive pressure and (either: "[[suck the cell on.|seal1]]","[[suck the cell on.|seal2]]")
(either: "Great", "Oh thank God", "Excellent", "Incredible"), it's (either: "rapidly", "quickly", "actually") [[sealing up|giga]].(either: "All right, it's sealing up, albeit slowly.", "The seal resistance increases just enough for you to know you didn't miss the cell entirely.", "It's not sealing up as fast as you'd like.")
You can (either: "[[wait patiently|wait1]]", "[[wait patiently|wait2]]") or (either: "[[suck more|suck1]]", "[[suck more|suck2]]")
(either: "It looks like you weren't close enough to the cell, because it's not sealing up.", "The square test pulse barely compresses. Were you even on the cell?", "The tip resistance rises to a pathetic 20 MOhm.")
(either: "Oh well, swing and a miss.", "For God's sake.")
[[Try again.|start]]You get absorbed in scrolling through your (either: "Twitter", "Facebook", "Instagram", "newsfeed", "junk mail") for a few minutes, but the seal hasn't improved at all.
(either: "Why hope at all?", "The world has no redeemable qualities.", "There is no God.", "You should've done an engineering degree.")
You decide it's time to [[start again|start]].You decide to use this as an opportunity to (either: "check your emails on your phone", "close your red, dry eyes for a moment", "put your head on the desk", "pray for mercy").
(either: "When you look back to your computer screen, you", "You then return to the computer to") see that the seal resistance has improved [[remarkably|giga]]. (either: "Yay!", "Good.", "Cool.")(either: "You decide to help the seal along by sucking a bit more", "You bring the mouthpiece back up to your lips and inhale slowly").
That turns out to be a mistake. (either: "A good portion of the cell enters the electrode", "The electrode moves and falls off the cell", "The entire cell is sucked into the electrode"). (either: "Really!?", "What? You didn't even suck that hard!", "Why did that happen?")
Time to [[start again.|start]]You give it a bit of gentle suction. The seal resistance starts rising more rapidly.
[[That's a good sign|giga]].The seal resistance is now over a gigaohm, so it's time to (either: "break in", "open it up", "go whole-cell").
(either: "With your finger over the hold switch", "Placing your finger on the zap button"), you (either: "[[give a sharp pulse of suction|break1]]", "[[give a sharp pulse of suction|break2]]", "[[give a sharp pulse of suction|break3]]")You give it a (either: "burst of suction", "short, sharp suck", "little kiss").
Your test pulse shudders and falls off the bottom of your screen. (either: "Whoops.", "Shit!", "Aaargh!", "Heck.")
[[Let's try again.|start]]You're in the cell, but (either: "it's not looking too happy", "it's taking a lot of holding current", "it's a bit shitty").
You can either (either: "[[wait for it to get better|breakwait1]]", "[[wait for it to get better|breakwait2]]") or (either: "[[just start recording|rec1]]", "[[just start recording|rec2]]").(transition: "pulse")[Success!] You're in the cell. (either: "But it's not over yet", "But this is just the beginning", "But you're not safe yet").
You note down the cell's passive properties and start your (either: "[[protocol|rec1]]", "[[protocol|rec2]]").You record (either: "a few", "a couple", "some") sweeps and then check your cell parameters again. The access resistance has crept up above what you want it to be. Your pipette is getting clogged. (either: "Sigh", "Jesus", "Why", "Come on, you little shit").
You can try to clear it either by [[gently|ra1]] [[sucking|ra2]] or [[carefully|ra2]] [[blowing|ra1]].You run all your protocols. The cell doesn't die, and the access resistance doesn't change.
(either: "Awesome!", "Sweet!", "Finally!") One data point!
(link:"Here's my actual website index.")[(gotoURL:"http://cocculin.neocities.org/home.html")]You wander away from the rig to pace around the lab for a bit.
(either: "The sudden exposure to bright light disorients you. You wonder to yourself if it's somehow unhealthy to be spending so much time in a dark corner", "You put some freshly washed glassware back in the cupboard and grumble to yourself about how the lab manager does nothing", "Some postdocs in the main labspace have mounted an inflated glove to a vortex and drawn a face on it. You laugh and take a video of it", "You check all the carbogen tanks in the lab to make sure that you're not going to be screwed over very soon").
When you reluctantly (either: "return", "come back to the rig", "get back to work"), you find that nothing has improved. Best to [[move on|start]].You watch your test pulse for a while, wondering if it's safe to leave it to (either: "use the bathroom for a second", "berate the lab manager for fun for a bit", "wander into the next room to chat with the people there", "grab a quick coffee", "make a cup of tea").
Just as you're about to go, it jumps up to an acceptable level of holding current.
Can't leave now, you need to [[start|rec2]] [[recording|rec1]].(either: "You happily see the capacitance transients perk up.", "The access resistance dips under twenty again.") (either: "Whew, that was tense.", "Thank God.", "Phew!")
Time to start (either: "[[recording|rec1]]", "[[recording|rec2]]").(either: "No, that was too hard!", "Shit!", "The test pulse plummets.") Your cell is (either: "dead", "done for", "toast").
(either: "You wish you'd joined a molecular biology lab instead", "You question whether life really is worth living", "You tried so hard, you got so far, but in the end, it doesn't even matter").
Oh well, [[try again.|start]](set: $s to (text-style: "shudder")) The electrode $s[won't stop shaking.]
You dropped (text-rotate:25)[it.]
There is a small dish of water. (click: "dish")[Your finger gets wet.]
A (either: "slimy slimy", "goopy", "slippery") puddle
<h3>Whole-Cell Patch Clamp Simulator V1.01</h3>
Click the [[purple text to start|start]].