Dear Valerie:
We study lipid absorption in the gut of Sparus aurata larvae, from mouth
opening (day3-4) to day 20.
We work on semi-thin (0.5 micron) and ultra-thin (90-150 nm) slides for
light and electronical (TEM) microscopy investigation respectively.
Here is the protocol we follow for our needs. After different trials it is
the one that gave us the best results in the study of liver (contrasting
and detection of lipids and glycogene) and intestine (contrasting and
lipids). It also very easy to proceed.
Larvae are anesthesied in cold sea water, prior to follow the protocol of
double fixation:
1 - glutaraldeid 2.5% in a cacodylate buffer 0.2M (450 nOsm) for 1 h
2 - rinsing in cacodylate 0.225M (4 times x 15 min)
3 - osmium tetroxide 1% in a cacodylate buffer 0.45M for 1 h
4 - rinsing in cacodylate 0.225M (4 times x 15 min)
5 - Dehydratation in alcohol of increasing degree (30°-10min; 50°-10min;
70°-10min; 95°-10min; 100°-3 times 5min)
6 - bath in alcohol 100° + Epon (1 vol : 1 vol) for 1 hour
7 - 2 baths in Epon (half a day each)
8 - inclusion
9 - polymerisation in a oven at 60° for 3 days and 3 nights
Hope this helps, and if you need, send me an Email to discuss the details.
alois
>Hi;
>I'm planning to do some histological study on the gut of Engraulididae
>larvae (around 6mm body length).
>Does anyone know what's the best way to preserve the larvae before the
>histological work?
>Secondly, has anyone done some work relating the
>length and morphology of Clupeids gut larvae to their alimentation?
>
>Thanks
>Valerie
>
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Alois Maurizi
Laboratoire de biologie animale (cc.102) ><> ><>
Université Montpellier II
34095 Montpellier - cedex 5 ><> ><>
France
Tel. ++33-04.67.14.36.77
Fax. ++33-04.67.14.36.75 ou 04.67.14.35.95 <><
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