Injection needle
Calibration: for 180 µm diameter egg, the volume is 3100 pl (3.1 nl)
0.25% injection (8 pl) is 2.5 div or 2.25 div
0.5% injection (15.5 pl) is 3.5 div or 3.5 div
1% injection (31 pl) is 4.75 div or 4.7 div
2% injection (62 pl) is 6.7 div or 6.25 div
3% injection (93 pl) is 7.25 div
So use
0.25% -- 2.5 div
0.5% -- 3.5 div
1% -- 4.75 div
2% -- 6.5 div
3% -- 7.25
SM-20
one turn of vertical knob is 50 µm. each marking is 1 µm
Focus through glass into air
have to adjust by 1 1/4 turns - move needle 62.5 µm further down in order to
get into focus
Light travels through 180 µm glass, and changes the focus level 62.5 further
down
Focus through glass and water
depends on how much water...
Typical "length" of oil cap = 225 µm. what volume is this?
to wash cover slips
100 ml in 200 ml tripour beaker?
heat 3 min microwave
add a pinch of alconox
drop half a box one at a time
let sit for 20 min
pour out, rinse well 5x
heat 3 min in microwave
pour out, rinse well 5x
drop one at a time into 1:1 dilution of concentrated HCl in glass bottle
let sit overnight
pour out, rinse well 5x tap water
rinse well 5x deionized water
drop one at a time into 50 ml centrifuge tube with 70% ethanol
loading capillary
to save RNA or whatever, I use smaller amounts
load a 20 ul pipetman with 3 ul silicon oil
put in a small amount of oil into capillary (about 0.7 ul?)
load 0.3-0.5 ul RNA in 10 ul pipetman
put this into the capillary - tricky, might not get in
put in another oil to finish
nothing at the back of the capillary
show picture
show picture of placement on injection slide, along with position of shelf
needle storage
how to make