MicrobeHunter.com Microscopy Forum

Hi all, in finger lakes/US we had a brief one week of below freezing temps. Good doggie and I had only to climb the hill to our : pond/stream mesocosm. Doggie not permited into this mesocosm as she might foolishly

plunge onto the ice of the pond segment ( pond 1/3rd 10 ft. depth, remainder 5 ft depth).

I was only able to scoop sample from one portion of the stream segment , an area which had open waters

Please enjoy this collection sample wet mount slide microscopy. When observations done ( perhaps two hours of microscopy enjoyment), I flushed the water bear, et all into my bench stable culture.

Thanks all for visiting this live wetmount slide microscopy enjoyment...microscopy world views of my neighborhood. charlie guevara/ finger lakes/US

#001 image: stream segment in foreground, pond in background.

Dinoflagellate wonderfully pigmented, and quite active..I chased this active winter protozoan for delightful and challenging image captures. Fresh water dinoflagellates are poorly studied in north america. images: 012, 025

Collotheca sp. of rotifer, in secreted gelatinous tube , a free living 'sessile rotifer' in our freshwaters. The long spines are moved to entrap small metazoan prey..push prey into that huge funnel mouth openning..that funnel mouth snaps shut.

This rotifer has a poorly ciliated corona.

Paramecium bursaria was quite irritated that I slurped it up and onto a wetmount slide.

Peritrich protozoa enjoy their algae thick matts. Some actually have endosymbiotic algae.

With such a narrow ( 'skinny') body form..I wonder how these peritrichs have room for daily heterozoic engulfment of foods..withthe dense cluster of algae within the protists body volume.

It's cool that you have such easy access to a lively source of microbiology!

Water bear graced my first wet mount slide, quite a microscopy observational treat.

Ahh, this crazy weather/ changing 'new normal' weather...by 1/26/24..all snow on ground melted..ice in the stream-pond meso cosum mostly melted. This worries me.

The area streams were all swollen with melt water..my good doggie sensibly did not enter the rapid angry stream flow.

Hi, squint a lot, thanks for looking. Any microscopist can collect area ( native assemblages) from their locals open spaces...and enjoy a vibrant microcosm..no need for a mesocosm as I fabricated.

I enjoy native fish , and i foolishly attempt to in my stream segment...establish freshwater mussels native to my finger lakes/US locale.

Wow but altered winters are brought home in finger lakes/NY. charlie guevara

Charlie G,
Always a real treat to see Good Doggie outside enjoying it all !

Yup, dennis, this breed is a 'winter dog'...in 'dog days of summer'..this breed is an: 'air-conditioner dog'.

We hail from Bergen county/Ridgewood, NJ...I hope your NJ winter has been going okay.

Many of these gold/orange colored dinoflagellates encountered with each wet mount slide I freshly pipet up from the now partially thawed ( much open waters) stream section of my mesocosm.

The wetmount slide also contained two water bears plodding along shafts of algae.

Most wonderful for me, a first time ever observation (for me a first time observation) of a curious rotifer behavior which I stumbled upon with my wet mount slide prep..for me it recalls circa 1879 Jean Henri Fabre's writings on observed

insect behaviors, specifically when noted educator, noted naturalist, noted writer Jean Henri Fabre observed, described in writing, illustrated, a collection of caterpillars marching around the rim of a tea cup..following the 'leader'.

On my second scoop water sample from the stream mesocosm ( after 1/27-1/28/24 abrupt ice thaw in our region), I noted a rotifer rapidly circling a small gas bubble...this rapid circling of the gas buuble..with the rotifers corona always

touching the gas bubble...went on for ten minutes. I fed huge globs of sample water to edge of this wetmount slide so I could attend to dinner preparations in kitchen, with out the slide drying up totally.

I returned to this wetmount slide...this rotifer still rapidly encircling the gas bubble, with it's corona touching the gas bubble.

I have never observed this behavior before...forum members perhaps may look for this freshwater rotifer behavior. the best to all, charlie guevara, fingerlakes/US

Hi all..about 2/22/24 temps plunged to 'normal winter' cold of: 19 degrees F. . Prior weeks were 40's-50's F....very troubling 'new normal weather' here in fingerlakes/NY..

I collected a scope sample from under the ice.

For me the excitement was my first encounter with peritrich ciliate epibionts attached in a dense cluster about a winter midge larva's mouth. This midge larva was quite healthy , powerfully thrashing about on my

wet mount slide. I can only speculate on how such an active winter midge larva was secured as a host by this dense cluster of stalked/ sessile ciliates..at it's mouth region!??

Today, 3/1/24..temps up to high 40's F. . charlie g

Under the 'winter ice' of this scoop collected sample...the wet mount slide first constructed ( strategic pipet slurps from...within surface algae matt, collection containers bottom sediments, scrape&slurp with the pipet along container wall)

contained a charming menagerie of quite irritated protists and meiofauna, Dinobryon colony of flagelates, a new to me rotifer, actively rotating on it's long axis as it rapidly swam about ( is this the same rotifer Genus I encountered

methodically swimming non stop about a gas bubble in my 1/24/24 wetmount slide?), a small heliozoan amoeba with large contractile vacuole on one side, and a water flea/Cyclops with no egg bags.

Please enjoy these 'last of our regions new normal winter' mesocosm neighbors of mine. charlie g./fingerlakes,US

Hi all, this early 3/24..only one day of snow, below 30 degree-F temps..with strong 50- 60 MPH winds. I scooped sampled an area of my stream segment of the mesocosm.

On the first wetmount slide prep I developed with pastuer pipet slups from the scoop sample jar...two (2!) Chlorohydra viridissima...two hydras randomly captured on the first wetmount slide..for me this is better than: 'bass fishing contests'!

One of these green hydras had eight (8) tentacles, the other green hydra had seven (7) tentacles...I am curious about this..much to online, and with textbooks look into this issueof: 'number of tentacles' in freshwater green hydras.

Although both these 'green hydra' are rather small in size ( relative sizes freshwater hydras exhibit )....I am wondering about this variation in number of tentacles in a mesocosm population of green hydras.

Please visit, please enjoy these finger lakes/US neighbors of mine. charlie g to image capture the entire hydra..I needed to use: 2X Plan objective. then I used : 4X plan objective.

Here ( image #024 ) is the eight (8) tentacle green hydra. Sure enough ( I always control water depth/ water film thickness under a wetmount slide cover slip..to not 'crush' meiofauna)...sure enough , this hydra is exploring and leaving

the water film under the coverslip. For me I fancy this a chance to 'touch base with', to shake a tiny hand..err..a tiny tentacle..of my freshwater meiofauna neighbor.

The number of algae ( Chlorella species) per hydra host cell stays constant in all individual green hydra, maintaining this constancy seems more difficult in the larger green hydra strains.

Green hydras seem to have evolved a dependence on Chlorella algae. These algae have probably not evolved a dependence on hydra.

Loeewenhoek first described hydra in 1702. In the 18th century Trembley ( Abraham Trembley; 1710-1784) collected and described green and brown hydra. Trembley established hydras remarkable capacity to recover from

damage that would kill most animals.

There existed in the 18th century a belief that animals at birth were endowed with a kind of uniqueness, an animal soul. Transforming a single hydra into ten (10) others identical with itself by slicing it ( the hydra) into pieces

effectively eliminated the idea of a unique animal soul.

source of my post: "Ecology and Classification of North American Freshwater Invertebrates", 2nd edition, 2001, James H. Thorp/ Alan P. Covich"..pgs: 145-147

Thanks Charlie, your reports are a welcome winter reprieve for this ice bound hobbyist. I’m still sorting through water collected last fall and pickings are getting slim.
Could you please provide a bit more information on what you use for slurping? I’ve tried drawing out plastic Pasteur pipettes with only moderate success. I’d like to try drawing out glass pipettes but wonder what size to start with.
And please keep up with the wonderful collection stories.
Harry

Hi, harry, thanks for visiting this thread. I always wondered about bitter cold Manitoba/Canada stream flows in harshest of winters...sure enough your Winnipeg/ Manitoba weather and news station stated from 12/23- to 2/24

Manitobans experienced near record warm 'winter weather'..of course with temp plunges for short spells for a few days..then back to 'warmer winter'. In winter season..stream flows continue under possible ice covers. due to ground water mix with stream flows..

ground water temps nearly constant year round ( so in dreadful summer heat..streams are indeed moderated by the mix into streams of ground water). Long story short..I thought your pond/stream microscopy could continue with a brisk

hammer blow to surface ice..to sample winter pond/stream microscopy assemblages..only, only if it is safe, you know the specific terrain ( shallow waters, no high stream banks, etc., etc..).

I as highschooler was excited about (with my own hands) drawing out glass pastuer pipetts to extremely fine 'micro-pipetts. Under the forgiving large working distance of 4X objectives..often 10X objectives ..to collect up individual protists and

meiofauna. But the rubber bulb action on a pipett drawn to a 'micro-tip' is highly erratic...the suction forces often splatter your target fluids high up the glass..even into the rubber bulb.

I now sense that 'micro-tip pipetts' are a tool if you need to isolate/ transfer one cell to a culture media for growth of a horde of the wee-beasties. And 'micro-tips' a pain to constantly flush series to keep this tool clean..and these tools

are oh so delicate to store between uses.

Stock 'right out of the box ' borosilicate glass pastuer pipetts are the tool to use..and these can be drawn 'micro pipet thin' over an alcohol lamp..or kitchen gas-range burner...I suggest you forget about 'micro-pipetts'..but with borosilicate glass

pipetts..you can make 'micro-tip' pipetts..or even finer 'glass wiskers' for manipulations on a microscope slide. All plastic 'disposable' pipetts..one with fine tip similar to glass pastuer pipetts I only use for 'pure clean chemicals'..not for

specimen fluids..I use these 'disposable' plastic pipetts dedicated to each chemical fluid I transfer with them..I reuse these for same chemiclal multiple times.

So here are my suggested protocols for bench development of wet-mount slides .

Going back and forth between the three bench images I posted..here are protocols I use to effectively 'round up' protists and meiofauna for constructing a wet mount slide.

By using large rectangular cover slips ( 22X40mm, No.1, globe scientific inc., Paramus, NJ)..you can safely observe in over 80% of the wet mount areas..avoiding the border coverslip areas..thus not muck up your higher-magnification/ tighter

working distance objectives. You also learn to feed your wetmount slide to not have a huge water column depth under the cover slip..yet never let this sandwich dry-out..or crush target organisms on the wet mount prep. If you in error 'over feed'

waters to your slide..a small piece of paper toweling can drink back excess waters from your wet mount. If you wick up excess waters from the slide from side opposite the side you added too much waters..well all the better..as you have

flushed entire slide with the fresh sample waters ( and new organisms).

A flat all white paper plate is quite a contrast enhancer when a specimen jar, or a sample poured into a dish , is placed on this white paper plate. You also place the field jar ( outdoors wet and perhaps some 'muck' on the container base) on

this plate right beside your scope..for 'specimen round ups'..keeps your bench tidy..resuse that paper plate for months..unless you really mess it up.

Use an LED torch to side illuminate either your specimen field container..or your specimen dish..enchances 'plunge and slup' with eye-dropper for reall fast/ reactive water fleas, worms, etc..

For the rest of target organism collecting..pastuer pipetts work fine. The bore of these pipetts about 1.0mm..this fine enough to collect target protists/ smaller meiofauna.

Scissors essential to snipe a length of filamentous algae..or other strands of plants..to an amount your slide can utilize ..after drape of strands on slide..tease it open..spread it well..fantastic communities reside within these plant

strands.

I use a glass platform for my slide, and for the rectangular cover slip..easy pick up of each from these raised platforms...of course if you are compiling a collection of target organisms for a wetmount slide..a second slide on it's own platform

needed...time to time drink off waters from the 'round up slides water droplet with target organisms..and continue your round up.

So, harry, eye-dropper bulb tool, and out of the box borosilicate pipetts the main collection tools. An elongated water sample droplet on a glass slide permits pastuer pipetts specimen collection 'naked eye'.

With 4X, or 10X objective..you observe a quarry organism..crisply slurp that region of sample with a pipett to a slide..observe that slide for the target.

Again I wish to state..it totally by chance that I encountered two green hydras on my first wetmount slide sample..of a container scoop sample of my mesocosm waters.

All the best, charlie g, finger lakes/US

Thanks so much Charlie, for this trove of wonderful information! I appreciate your taking the time to put this together.
Our winter here in Manitoba was mild but thick ice doesn’t really melt that fast. Ice typically gets over 3 feet thick and people around here are still happily ice fishing on it.
I had slightly modified your long water drip along the length of the slide to putting 5 or 6 drops instead. That way I had a better chance of isolating a specimen when found. When I used your long sample along the slide method I suspect any protists I tried to pick up ended up stuck in my plastic micro pipette.
I have a long glass pipette with rubber bulb but I find it pretty useless.
So later this morning I’m going to order some micro pipettes and perhaps some shorter eye dropper bulb tubes.
Also love the idea of a white paper plate and side lighting.
Almost forgot, I do use the longer cover slips already, especially when using an oil objective with a wet mount slide.
Did a quick search and found these. Do they look right?

https://www.amazon.ca/iplusmile-Glass-C ... 9hdGY&th=1

Thanks again and Happy Microbe Hunting!
Cheers, Harry

You can low cost purchase a rubber bulb activator for all sorts of capillary tubes...but my capillary tubes, my 10 and 20 microliter glass capillary tubes sit in a draw, unused. If you have trouble with your pastuer pipetts bulb action being

useless..blame the problem on 'crappy' rubber bulbs..those approximately 1.0 mm bore pastuer pipetts work fine with a well sealing rubber bulb...I would be lost without this tool kit.

Most importantly ( heh-heh, for our forum enjoyment...hint, hint, harry)..ask your ice-fishing buds to snagg some benthic plants, other structure..for your microscopy observations. If you yourself have access to an ice-fishing hole...

get going on collecting bottom structure!

When my son took me on a classic wooden boat(?1956 Chris-Craft wooden boat?) fishing charter on canadian side of St.Lawrence river this past 8/23, the captain sensed I was nuts..I was excited about the plants my line brought

between largemouth bass captures...we could only use earthworms, canadian side of river does not permit minnows as bait. The good captain gave me a large zip-close plastic bag to hold my river dredgeings in..but he thought I was nuts.

all the best, charlie g, fingerlakes/US

Thanks Charlie, will look that stuff up to see if its available in the land of the polar bear.
As far as ice fishing goes, age has made me wiser and I stay in the comfort of a warm house. Fishing is for spring, summer and fall.
We are nuts!
Harry

This is an epic! Congratulations.

charlie g wrote: ↑

Tue Jan 30, 2024 4:19 am

On my second scoop water sample from the stream mesocosm ( after 1/27-1/28/24 abrupt ice thaw in our region), I noted a rotifer rapidly circling a small gas bubble...this rapid circling of the gas buuble..with the rotifers corona always

touching the gas bubble...went on for ten minutes. I fed huge globs of sample water to edge of this wetmount slide so I could attend to dinner preparations in kitchen, with out the slide drying up totally.

I returned to this wetmount slide...this rotifer still rapidly encircling the gas bubble, with it's corona touching the gas bubble.

I have never observed this behavior before...forum members perhaps may look for this freshwater rotifer behavior. the best to all, charlie guevara, fingerlakes/US

Very interesting, Charlie G. I am guessing that might be because rotifers prefer to have ample oxygen and the air bubble provided oxygen.

Happy spring'24..first hydra bud of this year encountered 3/30/24. All surface ice in the mesocosm gone.

It was 12/25/1702 that Antoni van Leeuwenhoek sent a letter to the Royal Society in UK, describing observations of active hydras Antoni collected. I am deeply puzzled by the sparse facts stating Antoni hired an illustrator to

draw the wonderfully biologically accurate illustrations which Antoni included with his letters to the Royal Society. Did Antoni train the illustrator to use his one lens microscope, then did Antoni and illustrator swap back and forth the

microscope setup with hydras?!! The tedium of simple oral free narration of objects being observed...to an illustrator next to the microscopist seems too tedious. I'm sure a professional forensic illustrator in police departments could

offer insights to : 'how Antoni's illustrations were generated.'

In my encountered Chlorohydra species ( this hydra has eight tentacles)...it is quite clear the symbiotic endogenous Chlorellae algae reside in the inner gastrodermis cells, not in the surface epithelial cell layer.

Similar to my pleasure in feeding family and friends, and coworkers since teenage years..I respectfully observed this green hydra capture both a cladoceran water flea, and a large nematode.

Please go back to the higher magnified tentacle image and for yourself..see if the clusters of endosymbiotic Chlorella algae ( these algae are said to be contained within individual gastrodermis layer cells..thus the clusters of algae)...

please see if you can count number of algae cells per cluster...see if you sense: same number of algae per gastrodermis cell containing these happy algae. all the best, charlie g

Ooops, sorry I missed your kind visit to this thread, and thank you for your speculation,zzffnn.

All my wet mount slide preps have rotifers and entrapped air bubbles...I am observing the wet mount slides within minutes of 'dropping the cover slip' on the sample droplet. No other rotifers..either on this wet mount slide,

or other wet mount slides from this winter'23-24...no slides manifested a rotifer so entrained to a gas bubble. New to me, this rotifer frenetic/ constant swimming behavior. No other rotifer on the imaged slide manifested this activity.

This is why I thought of the great french naturalist, Jean Henri Fabre's report and illustrations of caterpillar's 'following the leader'...round and round on the lip of a tea cup...never stopping their march.

I hope some microscopist with rotifers on their observation list...I hope they encounter this swimming behavior. charlie g...getting ready for our 4/8/24 total solar eclipse!