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RuralAnomaly
Sporadic


Registered: 10/05/13
Posts: 2,153
Loc: Spitzenkörper Ohio
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measuring spores without a reticule
#19114114 - 11/10/13 10:18 AM (10 years, 2 months ago) |
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i've been wracking my brain for a few days (not a pretty thing if you're anywhere close)
primarily, how to compensate for less than ideal equipment microscopically, and still get useful information, esp. regarding spore dimensions.
here's my chain of thinking so far.
I got wind of piximeter from one of Alan's posts and i can muddle through the french a bit and even luckier, cuz i'm rusty, the program installs in english. so that's installed.
now to the part i'd like discussed: since some of the images you can find show the dimensions AND a reference bar ( |---------| equals 200 µm for example )
could you use THAT bar to create a reference against which other images that [i would] load into piximeter? first load the above image and use that as the calibration reference then, i image some spores from a macroscopically well id'd species and measure its spores (preferably even same species as above?)
its sort of going backwards, and may be convoluted and i know that largely the propagation of error is going to come from the variance in the spore size of the species i select to use as a means to calibrate MY scope against the literature (so perhaps ideally choosing one with as little variance in mature spores as possible to find) AND i just started messing with the program so its subtle niceties are lost on me as of yet, but it seems to be a v. useful tool, if not this way then definitely down the line.
there may be some serious flaws in my logic here due to some factors i haven't considered (either out of flat out ignorance that they are factors, or just me being generally ignorant) but wouldn't it be wunnerful if technology gave us yet another work-around to compensate for no reticule
TIA for your thoughts on this
oh and please mention any other pixel measuring software you use (esp. free/shareware lol)
-------------------- bite my basidiocarp
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Byrain

Registered: 01/07/10
Posts: 9,664
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Re: measuring spores without a reticule [Re: RuralAnomaly]
#19114194 - 11/10/13 10:36 AM (10 years, 2 months ago) |
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You calibrate your scope with a stage micrometer because its a known size, the reticle in a eye piece and any images you find online wont necessarily conform to those sizes.
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RuralAnomaly
Sporadic


Registered: 10/05/13
Posts: 2,153
Loc: Spitzenkörper Ohio
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Re: measuring spores without a reticule [Re: Byrain]
#19114266 - 11/10/13 10:51 AM (10 years, 2 months ago) |
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i guess the idea behind it is solving for unknowns using algebra or ratios whichever it is
just using made up numbers here to flesh out my thinking:
from reference pic if spore A = 10µm which in pixels=40 and its 'reference bar' of 100µm would then be 400 pixels (on my screen only i understand, i'm creating a measurement unique to my gear and might not be the same everywhere)
this i use as my calibration reference for piximeter
i generate my own picture of maybe preferably the same type spores.
now, in my homemade picture, those spores all measure 8µm using piximeter 'uncalibrated'. therefore, i'm 20% off vs the referent and would need to calculate accordingly for future measurements on MY rig of computer/camera/scope OR possibly be able to create an alternative 'calibration' using piximeter for subsequent photos.
math makes my brain hurt lol
does this make sense?
-------------------- bite my basidiocarp
Edited by RuralAnomaly (11/10/13 10:52 AM)
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