Smolke:Protocols/Guidelines - Revision history

Kathy Y. Wei: /* 96-Well Plates */

2012-11-18T21:48:16Z

96-Well Plates

←Older revision		Revision as of 21:48, 18 November 2012
Line 35:		Line 35:
	==96-Well Plates==		==96-Well Plates==
	*The VYB can only run ONE full 96-well plate if:		*The VYB can only run ONE full 96-well plate if:
-	1) It has a FULL bottle of running buffer and an EMPTY waste bottle.	+	# It has a FULL bottle of running buffer and an EMPTY waste bottle.
-	2) Flow rate is set to HIGH.	+	# Flow rate is set to HIGH.
-	3) Mode is set to SCREEN.	+	# Mode is set to SCREEN.
-	4) Sample volume run is set to <50 uL.	+	# Sample volume run is set to <50 uL.
	*The cooler for 96-plate is kept at the bottom shelf of deli fridge. -YHW		*The cooler for 96-plate is kept at the bottom shelf of deli fridge. -YHW
	*Make sure the running buffer bottle is full and waste bottle is empty before EACH 96-well plate.		*Make sure the running buffer bottle is full and waste bottle is empty before EACH 96-well plate.

Kathy Y. Wei: /* 96-Well Plates */

2012-11-18T21:45:04Z

96-Well Plates

←Older revision		Revision as of 21:45, 18 November 2012
Line 34:		Line 34:

	==96-Well Plates==		==96-Well Plates==
-	*The VYB can only run ONE full 96-well plate if it has a FULL bottle of running buffer and an EMPTY waste bottle.	+	*The VYB can only run ONE full 96-well plate if:
		+	1) It has a FULL bottle of running buffer and an EMPTY waste bottle.
		+	2) Flow rate is set to HIGH.
		+	3) Mode is set to SCREEN.
		+	4) Sample volume run is set to <50 uL.
	*The cooler for 96-plate is kept at the bottom shelf of deli fridge. -YHW		*The cooler for 96-plate is kept at the bottom shelf of deli fridge. -YHW
	*Make sure the running buffer bottle is full and waste bottle is empty before EACH 96-well plate.		*Make sure the running buffer bottle is full and waste bottle is empty before EACH 96-well plate.

Kathy Y. Wei: /* Calibration */

2012-07-23T21:28:10Z

Calibration

←Older revision		Revision as of 21:28, 23 July 2012
Line 23:		Line 23:
	*Follow the instructions on the screen.		*Follow the instructions on the screen.
	*If calibration does not pass:		*If calibration does not pass:
-	**Make sure the lasers have had >30 min to warm up.	+	**Make sure the lasers have had >30 min to warm up. If this is not the issue:
-	**Warm up 1% hypochlorite to 37oC, re-run with warm bleach, and re-run beads.	+	**Warm up 1% hypochlorite to 37oC, re-run with warm bleach, and re-run beads. If this does not work:
-	**Contact VYB support under 'Tools' tab > 'MACSQuant live support'.	+	***Contact VYB support under 'Tools' tab > 'MACSQuant live support'.
-	**Let Kathy know ASAP.	+	***Let Kathy know ASAP.

	==Shut Down==		==Shut Down==

Kathy Y. Wei: /* Calibration */

2012-07-23T21:27:06Z

Calibration

←Older revision		Revision as of 21:27, 23 July 2012
Line 25:		Line 25:
	**Make sure the lasers have had >30 min to warm up.		**Make sure the lasers have had >30 min to warm up.
	**Warm up 1% hypochlorite to 37oC, re-run with warm bleach, and re-run beads.		**Warm up 1% hypochlorite to 37oC, re-run with warm bleach, and re-run beads.
-	**Contact VYB support under Tools tab > ~~Remote~~ support.	+	**Contact VYB support under 'Tools' tab > 'MACSQuant live support'.
	**Let Kathy know ASAP.		**Let Kathy know ASAP.

Kathy Y. Wei: /* Start Up */

2012-07-23T21:08:22Z

Start Up

←Older revision		Revision as of 21:08, 23 July 2012
Line 14:		Line 14:
	*Run back flush (~16 min).		*Run back flush (~16 min).
	*Run clean with 1% hypochlorite (~10 min).		*Run clean with 1% hypochlorite (~10 min).
		+
		+	==Calibration==
		+	Calibrate the machine using the MACS Quant beads when the machine asks (notification at the bottom of the screen, near the left).
		+	*Put the machine in single tube holder mode.
		+	*Briefly vortex the beads to mix. Beads are in the fridge under the Biacore, on the bottom shelf.
		+	*Click the barcode icon near the top screen.
		+	*Scan the barcode on the bead bottle.
		+	*Follow the instructions on the screen.
		+	*If calibration does not pass:
		+	**Make sure the lasers have had >30 min to warm up.
		+	**Warm up 1% hypochlorite to 37oC, re-run with warm bleach, and re-run beads.
		+	**Contact VYB support under Tools tab > Remote support.
		+	**Let Kathy know ASAP.

	==Shut Down==		==Shut Down==

James V. Vowles at 16:41, 23 July 2012

2012-07-23T16:41:29Z

←Older revision		Revision as of 16:41, 23 July 2012
Line 3:		Line 3:

	If machine is off:		If machine is off:
-	*Turn on machine by touching to screen.	+	*Turn on machine by touching the screen.
	*Logon and put the machine into data acquisition mode.		*Logon and put the machine into data acquisition mode.
	*Run back flush (~16 min).		*Run back flush (~16 min).

Yen-Hsiang Wang: /* 96-Well Plates */

2012-07-20T00:42:34Z

96-Well Plates

←Older revision		Revision as of 00:42, 20 July 2012
Line 22:		Line 22:
	==96-Well Plates==		==96-Well Plates==
	*The VYB can only run ONE full 96-well plate if it has a FULL bottle of running buffer and an EMPTY waste bottle.		*The VYB can only run ONE full 96-well plate if it has a FULL bottle of running buffer and an EMPTY waste bottle.
		+	*The cooler for 96-plate is kept at the bottom shelf of deli fridge. -YHW
	*Make sure the running buffer bottle is full and waste bottle is empty before EACH 96-well plate.		*Make sure the running buffer bottle is full and waste bottle is empty before EACH 96-well plate.
	*NOTE: In the future, there will be larger buffer bottles & this may no longer be a problem.		*NOTE: In the future, there will be larger buffer bottles & this may no longer be a problem.

Yen-Hsiang Wang: /* Filtering Samples */

2012-07-20T00:40:40Z

Filtering Samples

←Older revision		Revision as of 00:40, 20 July 2012
Line 36:		Line 36:
	**Clean-up:		**Clean-up:
	***Mammalian samples: Soak in 10% bleach for about 1 day, then soak in water about 1 day, then dry and return sheets to bag.		***Mammalian samples: Soak in 10% bleach for about 1 day, then soak in water about 1 day, then dry and return sheets to bag.
-	***Yeast samples: Soak in water for a couple of days, then dry and return sheets to bag. (YHW: I usually spread ~~EtOH~~ on it while drying)	+	***Yeast samples: Soak in water for a couple of days, then dry and return sheets to bag. (YHW: I usually spread ethanol on it while drying)
	*Cell strainers (BD Falcon Cat. No.: 352340)		*Cell strainers (BD Falcon Cat. No.: 352340)
	**Not consistently stocked. Order if needed.		**Not consistently stocked. Order if needed.

Yen-Hsiang Wang: /* Filtering Samples */

2012-07-20T00:40:23Z

Filtering Samples

←Older revision		Revision as of 00:40, 20 July 2012
Line 36:		Line 36:
	**Clean-up:		**Clean-up:
	***Mammalian samples: Soak in 10% bleach for about 1 day, then soak in water about 1 day, then dry and return sheets to bag.		***Mammalian samples: Soak in 10% bleach for about 1 day, then soak in water about 1 day, then dry and return sheets to bag.
-	***Yeast samples: Soak in water for a couple of days, then dry and return sheets to bag.	+	***Yeast samples: Soak in water for a couple of days, then dry and return sheets to bag. (YHW: I usually spread EtOH on it while drying)
	*Cell strainers (BD Falcon Cat. No.: 352340)		*Cell strainers (BD Falcon Cat. No.: 352340)
	**Not consistently stocked. Order if needed.		**Not consistently stocked. Order if needed.

Kathy Y. Wei: New page: ==Start Up== Instructions for machine start up and various other functions are on the lamenated guides near the VYB. If machine is off: Turn on machine by touching to screen. Logon and ...

2012-07-19T23:55:50Z

New page: ==Start Up== Instructions for machine start up and various other functions are on the lamenated guides near the VYB. If machine is off: *Turn on machine by touching to screen. *Logon and ...

New page

==Start Up==
Instructions for machine start up and various other functions are on the lamenated guides near the VYB.

If machine is off:
*Turn on machine by touching to screen.
*Logon and put the machine into data acquisition mode.
*Run back flush (~16 min).
*Run clean with 1% hypochlorite (~10 min).
**NOTE: 100% bleach = 15% hypochlorite.
**NOTE: Clean only runs if machine is in the single tube mode.
*Run calibration beads.

If machine is already on, assume it is not clean:
*Run back flush (~16 min).
*Run clean with 1% hypochlorite (~10 min).

==Shut Down==
*When finished, ALWAYS make sure there is ENOUGH BUFFER and ENOUGH SPACE in the waste container. Otherwise, shut down will stall and leave sticky, dead, clog-inducing samples in the machine until someone notices.
*Shut down the machine unless another user is signed up within a couple of hours after you finish, but check buffer and waste in case later user cancels.
*Machine will automatically attempt to shut down ~2 hrs after its last fluidics function. However, you must still check buffer and waste before leaving the machine.

==96-Well Plates==
*The VYB can only run ONE full 96-well plate if it has a FULL bottle of running buffer and an EMPTY waste bottle.
*Make sure the running buffer bottle is full and waste bottle is empty before EACH 96-well plate.
*NOTE: In the future, there will be larger buffer bottles & this may no longer be a problem.

==Empty Buffer Bottles==
*Rinse with water.
*Remove the label and lid.
*Recycle (blue atrium bin).

==Filtering Samples==
ALWAYS FILTER YOUR SAMPLES.
*Nylon filter sheets (http://www.amazon.com/Microns-Square-Opening-Thread-Diameter/dp/B000FN0OG2/ref=pd_sbs_indust_5)
**Available in zip-lock bag above VYB bench.
**Clean-up:
***Mammalian samples: Soak in 10% bleach for about 1 day, then soak in water about 1 day, then dry and return sheets to bag.
***Yeast samples: Soak in water for a couple of days, then dry and return sheets to bag.
*Cell strainers (BD Falcon Cat. No.: 352340)
**Not consistently stocked. Order if needed.
**Disposable.

==Quirks==
*The software is frozen.
**If you are running a sample, leave the machine alone. It is probably busy recording a lot of data and will recover once it finishes (up to ~15 min).
*I can't run 'Clean'.
**Make sure the machine is set in the single tube mode.
*My samples look weird in FlowJo.
**In FlowJo, adjust the file properties to match the decade settings used on the VYB.

Smolke:Protocols/Guidelines - Revision history

Kathy Y. Wei: /* 96-Well Plates */

Kathy Y. Wei: /* 96-Well Plates */

Kathy Y. Wei: /* Calibration */

Kathy Y. Wei: /* Calibration */

Kathy Y. Wei: /* Start Up */

James V. Vowles at 16:41, 23 July 2012

Yen-Hsiang Wang: /* 96-Well Plates */

Yen-Hsiang Wang: /* Filtering Samples */

Yen-Hsiang Wang: /* Filtering Samples */

Kathy Y. Wei: New page: ==Start Up== Instructions for machine start up and various other functions are on the lamenated guides near the VYB. If machine is off: *Turn on machine by touching to screen. *Logon and ...

Kathy Y. Wei: New page: ==Start Up== Instructions for machine start up and various other functions are on the lamenated guides near the VYB. If machine is off: Turn on machine by touching to screen. Logon and ...