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06-12-2018 07:35

Ethan Crenson

Hello all, These were found by my friend recently

08-12-2018 15:13

Per Marstad Per Marstad

This was growing on Fagus, 2-3 mm. The spores are

14-11-2018 19:07

Andgelo Mombert

Bonsoir,Sur bois mort de feuillu. Apothécies sess

09-12-2018 23:06

Joop van der Lee

Found on cow dung. Fruitbody is covered with tufs

06-12-2018 00:02

Elsa Sousa

Good evening,I need your sharp eyes on this ascos.

11-12-2018 17:31

Peter Püwert Peter Püwert

Can somebody help ? Massee G. et Salmon E. S. (19

11-12-2018 23:18

Iglesias Plácido

Hola amigos, est sortie la revue Errotari de cette

10-12-2018 23:47

BERNARD CLESSE BERNARD CLESSE

Que pensez-vous de cette récolte, faite sur crott

07-12-2018 17:24

Enrique Rubio Enrique Rubio

Quelqu'un pourrait-il avoir une copie de ce docume

10-12-2018 12:00

Tanja Böhning Tanja Böhning

Bonjour,   Hier nous avons trouvé encore une p

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Proper staining method?
Viktorie Halasu, 16-11-2018 19:57
Viktorie HalasuHello forum,

I'm looking for some protocol how to stain fungal tissues, specifically lipids with Sudan III. The biggest problem I have is how to remove excessive lipids from damaged spores and other structures that got spilled into the preparation, and yet preserve undamaged the lipids inside cells (spores and other). I tried rinsing it with 60% ethanol, but the lipids merely coagulated into bigger drops and stayed attached to the surface of the section. Using a kitchen detergent helped more but that one doesn't exactly look like a publishable method. : )
Thank you in advance.

Viktorie
Hans-Otto Baral, 16-11-2018 22:06
Hans-Otto Baral
Re : Proper staining method?
Not sure if I understand you well. I assume you mean that staining lipids requires damaged cell walls?

I can  actually confirm this for spores with thick walls. Only  a few times I tried Sudan and also found alkalinized Cresyl Blue (CRBA) useful. The following text I wrote in my VT paper in 1992:


Lipophile dyes are commonly used to give more or less specific stains for fatty matters (see KIRK, 1966: 87ff.). Most of these tests are lethal to the cells. I have tested Sudan III in lactic acid in se¬veral species: a distinct reddish stain of the LBs was rapidly obtained especially after heating the slide, while CRBA gave the characteristic amber stain. KIRK (l.c.) recommended two tests (using benzapyrene-caffeine, and neutral red) for staining LBs in statu vivo by fluores¬cence.
KORF & ERB (1972) and KORF (1977) found Trichophaeopsis bicuspis to differ from Trichophaea by ascospores with "non-oleaginous, somewhat resinous inclusions" instead of LBs since the inclusions failed to absorb oil stains such as Sudan IV. My material of T. bicuspis showed two polar LBs 3.5-4 µm in size. Indeed, the whole spore content remained unstained by CRBA or Sudan III for hours, even after boiling, except for immature, and ruptured mature spores, where the LBs stained amber in CRBA and red in Sudan III. Thus, the negati¬ve result is clearly due to the impermeability of the thick wall of the mature spore to these dyes.

Zotto
Viktorie Halasu, 16-11-2018 22:29
Viktorie Halasu
Re : Proper staining method?
Hello Zotto,
no, I meant that after some spores are accidently badly damaged (with the razorblade or by whatever cause), their contents will escape and float in the preparate. Now I need to observe some small lipid bodies (?) in cells of outer excipulum (it's dead herbarium material). But because of those cracked spores etc., I cannot be sure whether the guttule I'm observing is actually inside of the cell (= a morpholog. character), or if it is one of these freely floating guttules that's just sticking to the cell surface from outside (= an artefact). And these floating lipid particles I want to remove. I appologize I don't have an image at hand. 

Viktorie
Hans-Otto Baral, 17-11-2018 08:09
Hans-Otto Baral
Re : Proper staining method?
Yes, now I understand. The same problem I know personally. It is usually a matter of the living vs. dead state, that one cannot be sure if the guttules are cell contents or have a secondary, alien origin. Are the excipular cell walls impermeable for Sudan? I have little experience with that stain. With CRB it works, it esily penetrates the cell wall and it stains oil in a rose colour while vacuoles in  blue.