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InvisiblebodhisattaMDiscordReddit
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Spores Are For Microscopy Only! * 50
    #21365919 - 03/05/15 02:31 PM (6 years, 8 months ago)



I took a medium sized medium darkness cubensis print. it's a little over an inch in diameter.

I was going to try to find out how many spores were on this print, however when I got the whole print in 10mL of water I couldn't break all the clumps up, I tried using some glycerine and an ultrasonic cleaner and I still had quite a bit of clumps, this means I wouldn't get an accurate number if I counted on a counting slide. so I did this instead



I took a very small speck as you can see in the middle of the coverslip on the slide.


this is at 100x magnification brightfield (10x eyepiece and 10x objective)


this is at 400x brightfield (10x eye 40x obj)


this is the top left still at 400x


this is part of the top left at 1000x oil immersion brightfield(10x 100x obj)

that's just one speck from a print, easily 1/1000th of a print and that one single speck could make a 10mL spore syringe easily.

this miniscule speck easily has 10,000,000 spores
if you put that one single speck into a syringe each mL would have 1 million spores.
One million spores per PF cake.
this is why a little goes a long way with spores.

Most vendor syringes could be diluted 10 fold and still work just fine.

After doing all of this I took the cover slip and smooshed out the spores a bit


here at 400x you can a small grouping of spores


and here's some of them at 1000x oil immersion.

if you were to use that single print to make 10 syringes. each syringe would have at least a few hundred million spores and that's conservative.


pulled a thread of rhizomorphic growth from agar and put it on a slide. as you can see there's quite a lot going on in even a small little sliver. here's the elusive clamp connection
More pictures of mycelium and agar plates, a continuation of what you see here

Like what you see? give a +5 rating and I'll give you one back if I haven't already done so.
:cheers:


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Edited by Trusted cuItivator (03/26/15 07:50 PM)


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Offlinebw86
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Re: Spores Are For Microscopy Only! [Re: bodhisatta] * 2
    #21365939 - 03/05/15 02:34 PM (6 years, 8 months ago)

I know you weren't really expecting to count spores:cool: and this post is to show people how little of a spore solution they need :yesnod: right?


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OfflineItsScience
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Re: Spores Are For Microscopy Only! [Re: bodhisatta] * 1
    #21365948 - 03/05/15 02:37 PM (6 years, 8 months ago)

Super Cool!  They always say that even if you don't see a super dark print you have millions even in the most invisible part, this just confirms that.  Really cool for those of us w/o microscopes!


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Re: Spores Are For Microscopy Only! [Re: bw86] * 1
    #21365950 - 03/05/15 02:38 PM (6 years, 8 months ago)

damn, nice! a couple  hundred millions should be enough to start a grow or 2! :wink:


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Re: Spores Are For Microscopy Only! [Re: spacechildo] * 1
    #21365955 - 03/05/15 02:39 PM (6 years, 8 months ago)

Nice post!


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InvisiblebodhisattaMDiscordReddit
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Re: Spores Are For Microscopy Only! [Re: bw86] * 1
    #21365959 - 03/05/15 02:40 PM (6 years, 8 months ago)

Quote:

bw86 said:
I know you weren't really expecting to count spores:cool: and this post is to show people how little of a spore solution they need :yesnod: right?



I was going to count them but I couldn't get every little clump to break up, I was going to use a haemacytometer but with clumps it would be horribly inaccurate.


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Re: Spores Are For Microscopy Only! [Re: ItsScience] * 1
    #21365960 - 03/05/15 02:40 PM (6 years, 8 months ago)

Ballin'


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InvisiblebodhisattaMDiscordReddit
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Re: Spores Are For Microscopy Only! [Re: bodhisatta] * 2
    #21366015 - 03/05/15 02:54 PM (6 years, 8 months ago)

Quote:

bodhisatta said:
Quote:

bw86 said:
I know you weren't really expecting to count spores:cool: and this post is to show people how little of a spore solution they need :yesnod: right?



I was going to count them but I couldn't get every little clump to break up, I was going to use a haemacytometer but with clumps it would be horribly inaccurate.





speaking of


here's yeast on a counting slide I took a picture of



yeast and cube spores are about the same size so counting them wouldn't have been a problem if I could have got it to homogenize and break up all the clumps


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Edited by Trusted cuItivator (03/05/15 03:20 PM)


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OfflineMonty514
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Re: Spores Are For Microscopy Only! [Re: Sagescruffy] * 1
    #21366048 - 03/05/15 03:02 PM (6 years, 8 months ago)

This is awesome. The pictures are awesome. You are awesome.


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Re: Spores Are For Microscopy Only! [Re: Monty514] * 1
    #21366067 - 03/05/15 03:06 PM (6 years, 8 months ago)

Good stuff! :hellyeah:


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Offlinebw86
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Re: Spores Are For Microscopy Only! [Re: Monty514] * 1
    #21366068 - 03/05/15 03:06 PM (6 years, 8 months ago)

so you are saying yes you were going to try and count all the spores in a spore print?
I understand using haemocytometer but that's a square mm. AN entire print would take for ever to count.


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InvisiblebodhisattaMDiscordReddit
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Re: Spores Are For Microscopy Only! [Re: bw86] * 1
    #21366075 - 03/05/15 03:08 PM (6 years, 8 months ago)

Quote:

bw86 said:
so you are saying yes you were going to try and count all the spores in a spore print?
I understand using haemocytometer but that's a square mm. AN entire print would take for ever to count.



that's not how it works.


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Offlinebw86
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Re: Spores Are For Microscopy Only! [Re: bodhisatta] * 1
    #21366096 - 03/05/15 03:13 PM (6 years, 8 months ago)

bro there is no way you were gona count an entire spore sprint.Thank you for the useful pictures you posted to help people understand...

every spore print is a different size anyway and drop different amounts you can easily get 4 prints from one cap.
im not here to argue, i thanked you multiple times, i like you as a person and don't want to start some petty feud


Edited by bw86 (03/05/15 03:19 PM)


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OfflineChem-4 OG
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Re: Spores Are For Microscopy Only! [Re: bodhisatta] * 1
    #21366098 - 03/05/15 03:13 PM (6 years, 8 months ago)

Quote:

bodhisatta said:


I took a medium sized medium darkness cubensis print. it's a little over an inch in diameter.

I was going to try to find out how many spores were on a single print, however when I got the whole print in 10mL of water I couldn't break all the clumps up, I tried using some glycerine and an ultrasonic cleaner and I still had quite a bit of clumps, this means I wouldn't get an accurate number if I counted on a counting slide. so I did this instead



I took a very small speck as you can see in the middle of the coverslip on the slide.


this is at 100x magnification brightfield (10x eyepiece and 10x objective)


this is at 400x brightfield (10x eye 40x obj)


this is the top left still at 400x


this is part of the top left at 1000x oil immersion brightfield(10x 100x obj)

that's just one speck from a print, easily 1/1000th of a print and that one single speck could make a 10mL spore syringe easily.

this miniscule speck easily has 10,000,000 spores
if you put that one single speck into a syringe each mL would have 1 million spores.
One million spores per PF cake.
this is why a little goes a long way with spores.

Most vendor syringes could be diluted 10 fold and still work just fine.

After doing all of this I took the cover slip and smooshed out the spores a bit


here at 400x you can a small grouping of spores


and here's some of them at 1000x oil immersion.

if you were to use that single print to make 10 syringes. each syringe would have at least a few hundred million spores and that's conservative.

Like what you see? give a +5 rating and I'll give you one back if I haven't already done so.
:cheers:




Since I can't afford a microscope of my own yet, I very much appreciate these photos and I will be making note of them.

I hope to see more of these photos with your interpretation of whats going on. :bow2:


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Re: Spores Are For Microscopy Only! [Re: bodhisatta] * 1
    #21366100 - 03/05/15 03:15 PM (6 years, 8 months ago)

GREAT STUFF. a microscope is in my future for sure!


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InvisiblebodhisattaMDiscordReddit
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Re: Spores Are For Microscopy Only! [Re: Hashish] * 1
    #21366111 - 03/05/15 03:18 PM (6 years, 8 months ago)

just for you BW

Quote:

bw86 said:
bro there is no way you were gona count an entire spore sprint.Thank you for the useful pictures you posted to help people understand...

every spore print is a different size anyway and drop different amounts you can easily get 4 prints from one cap.
im not here to argue, i thanked you multiple times, i like you as a person and don't want to start some petty feud




I'm not starting a feud you're just getting confused because you don't know how a counting slide works. yes you can count the whole print, no you don't physically count the whole print. and yes every print is different, I was just counting MY print as an example. it's only representative of itself not any other print unless that other print is the same size just as dark and even then...


here's yeast on a counting slide I took a picture of

I counted 48 viable yeast cells on that middle square there.
that's 1/25th of 1x10-4thml

the yeast sample I put on the counting slide was diluted 1:100
IE 1ml yeast slurry into 9ml of water, shake that up. take 1ml of that solution and put it into 9ml of water. then take some of that and put it on the counting slip. now the yeast you're looking at is diluted 1:100 if you didn't dilute it there would be too many cells to count on the grid. you want between 20-100 cells on each of the squares so you can accurately count them.

so if I got 48 viable cells * 25(cus there's 24 more other squares I didn't count) * 100(dilution factor) * 104th to get back to mL

you get 1.2x109thcells per ml

for example say I scraped that whole entire print into 500ml of water.
I put it on the counting slide and I counted lets just say 50 spores in one of those squares. then I would know that there was ~6.25 trillion spores on that whole print. I couldn't get the spores to un-clump. so I couldn't put them on the counting slide. thus I don't know how many spores are on that print. I'll try it again some other day but there's more than trillions of spores on the print. I would guess 1x1010th as a conservative answer to how many spores are there on the whole print.


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Edited by Trusted cuItivator (03/05/15 03:23 PM)


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Re: Spores Are For Microscopy Only! [Re: bodhisatta] * 1
    #21366121 - 03/05/15 03:21 PM (6 years, 8 months ago)

Errr math  :trippingsheens:


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Offlinebw86
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Re: Spores Are For Microscopy Only! [Re: bodhisatta] * 1
    #21366123 - 03/05/15 03:21 PM (6 years, 8 months ago)

thank you :hatsoff:
but that would still be how  many spores dropped from that print.
not how many spores are in the print, or even an estimate to how many spores are in cube print in general.
i guess im just arguing semantics because you said how"many spores were on a single print" not that print.
:hatsoff: thank you for your contribution.


Edited by bw86 (03/05/15 03:27 PM)


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Re: Spores Are For Microscopy Only! [Re: bw86] * 1
    #21366163 - 03/05/15 03:29 PM (6 years, 8 months ago)

Quote:

bw86 said:
thank you :hatsoff:



so you see I'm not counting the whole print spore by spore. I'm counting a representative part of the print. I put the whole entire print into 500ml of water. then I take a fraction of a drop onto the counting slide. a counting slide works by using a special cover slip only a set volume of liquid is in the space between the cover slip and the slide so you have a known volume in which you're counting cells or spores. you count a small volume then multiply it by your dilution and then multiply it by the volume of the counting slide.

sort of like I take every single person on the planet and load them into the pacific ocean. I take 1000 gallons of pacific ocean and find 5 humans in there. then I multiply that up by the whole volume of the ocean and get an approximate number of people on the whole planet. this is why the sample has to be extremely homogeneous with no clumps.


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Re: Spores Are For Microscopy Only! [Re: bw86] * 1
    #21366176 - 03/05/15 03:32 PM (6 years, 8 months ago)

Quote:

bw86 said:
thank you :hatsoff:
but that would still be how  many spores dropped from that print.
not how many spores are in the print, or even an estimate to how many spores are in cube print in general.
i guess im just arguing semantics because you said how"many spores were on a single print" not that print.
:hatsoff: thank you for your contribution.



I changed the wording of the first post. I was going to count the spores on my print, I scraped the whole entire print off so there was nothing left on the foil. I put the print into 10ml of water then poured that 10ml into a graduated cylinder and brought it up to 500ml (50 syringes for one print basically)

but when I put the sample on the counting slide I could only see clumps of spores not homogenous so I gave up trying to count them.


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