What tubing should I order for plastic-capillary (SCORE) imaging?
The tubing is from Cole Parmer. One part is short sections of tubing
that are relatively straight. The other is a large roll, which you will
have to straighten yourself before using. Otherwise, it'll pop off the
top cover slip when you turn your specimen. The products are different
prices, so choose which is best for your situation.
EW-00244-ZU Tubing, FEP, 1/32" ID 1/16" OD, 450' - 431 USD
WU-06406-60 Tubing, FEP, 1/32" ID 1/16" OD, 25' - 14 USD
We use a 2.5% solution of embryo water with methyl cellulose. You have
to dissolve it in hot water. When it cools, add the usual concentration
of eugenol or tricane. Place a blob of the thick solution on a dish and
transfer the fry (with as little water as possible) into it. Mix gently,
then draw the embryos into the capillary, head first, using suction.
Draw in 2 cm of the thick fluid before the first fish. Space them out
a bit. After a few tries, you'll never go back to the depression slide!
What filter set should I be using for the GBT lines?
The filter set we use is P/N 489063. The excitation filter is a
band-pass filter centered at 572 nm with a half-power width of 25 nm.
The dichroic mirror reflects light with wavelengths below 590 nm and
allows higher wavelengths to pass through. The emission filter is a
band-pass filter centered on 629 nm, with half-power width of 62 nm.
In short, BP572/25, DICHROIC FT 590, EM BP 629/62.
Chroma has a similar filter set that should also work, although we
have not personally tested it. Their mCherry filter set is P/N 49008,
which has the following characteristics: Ex ET560/40x, Dichroic T585LP,
See the spectrum of mRFP1 below.