TIRF Microscopy

'Preparing Acid Washed Glass Cover slips (Salmon lab)'

This helps cells and polyamino acids stick to glass. It also helps your DIC pictures look prettier by getting rid of small particles stuck on the glass.

First separate cover slips  from one another and place them into a glass container with a lid containing the amount of water you'll need to add in order to make a 1M HCl solution. It is important that the cover slips are not sticking to each other because you will need to use one at a time and it's easier to do this now rather than later.

Be careful when changing solutions not to pour out cover slips into the sink which can both block the drain and be really irritating to some lab members. If you do this, then simply clean them up (be sure to rinse the sink well first if you spill HCl along with the cover slips.

1) Heat cover slips in a loosely covered glass beaker in 1M HCl at 50-60oC for 4-16h.

2) Cool to room temperature

3) Rinse out 1M HCl with ddH2O

4) Fill container with ddH2O and sonicate in water bath for 30 mins

5) Repeat step 4

6) Repeat step 4

7) Fill container with 50% EtOH and 50% ddH2O and sonicate in water bath for 30 min

8) Fill container with 70% EtOH and 30% ddH2O and sonicate in water bath for 30 min

9) Fill container with 95% EtOH and sonicate in water bath for 30 min

10) Fill container with 95% EtOH

Preparing Chambers

Materials: tapes, acid washed coverslips immersed in ethanol, Blade, burner, tweezers, slides

to almost equally divided tapes
 * 1) Cut tapes with each length of 3-4cm. Use a slide to prepare tapes, use tip of blades

0.5cm gap
 * 1) Take one cut tape and stick onto another slide, doing so twice with each tape in a

kimwipe
 * 1) Use  tweezer grasp a coverslip and then remove excess ethanol on it with

get a small flame and then gone)
 * 1) Heat coverslip with tweezer for a bit over burner to remove all ethanol (will


 * 1) Stick coverslip onto slide in the center of 2 tapes


 * 1) Use tweezer to press the coverslip to ensure it is secured

Arrange all slides in a box
 * 1) Heat the slide for a bit over burner to melt and seal tape