Cyclopian garment change...

Cyclops reluctantly assumed a side pose for the camera

Spring has made a tentative approach but is in steady retreat again...temps are heading down to freezing again later today. The warm weather of the last few days did however open a ditch along my property and I've finally got a whole bunch of stuff to examine. Cyclops, ostracods, and a whole bunch of different ciliates with many rotifers among them. My technique, if I can call it that, was to open a jar at the bottom of the ditch, right above the sediment. Doing this allowed a bunch of water to rush into the container bringing everything within a few inches into it, including some debris. back at the microscope, one micro-event did stand out. As I was examining one of the copepods, it started to molt and in just a few seconds it was out of it's old skin and motoring away. Luckily, I had been taking pictures and I got a couple of, well, mediocre images that show what happened. I wonder if the event was serendipitous or the molt was brought on by the pressure of the cover glass?

Bubble on upper back shows where old skin has started bunching up

Discarded skin on left, newly clothed cyclops on the right

Books that instruct and entertain...me anyway

This book, published in 1960, will fit right in with those of us using microscope models sold during that era. It is based on a course delivered by the author and designed for students and hobbyists. Is there anyone reading this blog who doesn't fall into one of those categories?

 A great contemporary book for those interested in taking pictures through the microscope. The author understands his subject and has done a good job of transcribing that knowledge to the pages of this book. He is also known to frequent one of the Yahoo microscope groups where he helps with questions and posts about his hobby.

 The best book I've found to help decipher the identity of those unknown critters found in many drops of water. It's set up as a key for protozoa with ample photos and drawings to help with the ID.

 Another great book, and the title says it all. A mixture of technical information and essays on observations that will be both helpful and enjoyable. The book is no longer in print and demand has pushed the price of used copies far beyond the original price. I love what the internet has done to book accessibility but hate what it's done to prices.

This is an eBook about a comprehensive protozoan study in a small bog area of Germany. The book is filled with photos and is a
available online for a reasonable amount.

















And then there is a whole treasure of books that are no longer copyrighted and freely available for download. Many of these are extremely relevant to what we're doing at the microscope today, in addition to being well written in a somewhat archaic tone. Most are littered with hand drawn illustartions. Here are three that I'm reading right now.

 Common Objects of Microscopes

 Evenings at the Microscope.

 Half Hours with the Microscope.

DIY scanning microscope...

I've tried using my Zeiss Universal to scan for specimens in a petri dish but the stage was NOT designed for that. After flipping the dish onto the floor once and other times balancing it while trying to stay in focus I had had enough. Yesterday, while mixing a cool beverage with watching my wife rake the lawn, the fermenting idea coalesced into a solid plan. After supper I hit the machine shop and started milling a couple of discs; one out of aluminum, the other plastic. Held together with some thin, double-sided tape and a couple of screws, it fit perfectly into the existing stage housing. And I was now able to slide around a petri dish to my heart's content, just like the old days with my student microscope.











 Due to lack of clarity on my part, I've been asked for additional photos to explain my device. In response, I offer the following:

Here is a photo of the original stage in place, complete with all paraphernalia that allows manipulating a slide using geared mechanical movement in the X and Y direction. Unfortunately, it is less friendly when trying to scan a petri dish.

Pictured here is the stage base, with stage removed. The three contact points are evident; two in the back and a spring-loaded one at the front. The new flat stage is designed to lock into these same three contact points.

And here is the finished product, a flat, unobstructed surface. The petri dish is moved around until a victim is isolated under the objective. The water, including said victim, is then drawn up into a pipette and deposited elsewhere in the dish or onto a slide .

Mite I find an adult or another egg?

 The mite of the previous post had piqued my interest and I was anxious to learn more. A search of the internet failed to provide any new information so it was back to the sample for either an adult or another egg. No adults but I did find another egg, this one intact and still containing what appears to be a fully formed specimen. The shot to the left is focused on the hairy fringe which seems to cover the whole egg. Four legs are evident near the bottom of the egg and a dark area might indicate where the head is located.

Here is another area of focus, this time nearer to the upper, outer surface.

At this point something odd happened that I just didn't catch while viewing through the microscope. Either some debris floated up to the egg or the evaporating water brought some debris closer to the egg. In either case, the bottom-right area of the egg was covered with small living organisms, as shown in the image to the right. There were two sizes of circular visitors, the small ones easily seen in the image and some much smaller. No idea what they are but the smaller ones might have been bacteria size. Or did the pressure of the cover slip due to receding water cause the egg to break?

Here is a darkfield shot of the scene showing the general mayhem surrounding the egg. The whole field to the bottom right of the egg is a mass of moving things, for want of a better word. I wish I had recorded more of what happened and other details like magnification.

A mite fuzzy...

I ran into an interesting experience yesterday while strolling down the microbial highway. There, not more than 10 inches away, I saw what appeared to be a new born mite struggling to get out of its egg case. Camera always at the ready, I snapped a photo so the scene details could be shared with you. The emerging mite appeared to be about 30 microns wide.

A few things were working against me. This egg, if that’s what it actually was, came from some tree lichen so the little beasty may have been terrestrial in nature. Since it was immersed in water as part of my plan to capture the elusive water bear, it may have been experiencing some distress. I was also having trouble with image focus; perhaps the mite was too thick or opaque for proper focus or illumination. It also seemed to be completely covered with a fairly thick layer of bacteria, caught in the image as a lighter halo surrounding the emerging mite and egg . When the shutter closed, only the upper two legs seemed to be in focus. So there we have it...poor picture but an interesting event I wanted to share. 

After this was all over the little critter was transferred to a drier piece of moss in the hope it may successfully continue its passage to eventual adulthood.

Another visit with the 8 form test diatoms...

This feels like self-flagellation given the state of my camera setup but I just had to see what my 100X objective would do with these diatoms. I had read a number of articles on this exact subject on the Microscopy-UK website and the results others were getting were, to say the least, tantalizing. Since I normally use the 0.63 NA condenser, I removed it and put in a 1.40 NA condenser.  A drop of immersion oil on the condenser, lower the slide onto it, another drop on the slide, move the 100X/1.30 oil objective onto the slide and I was away. The filter tray had a green filter and the condenser and aperture diaphragm were adjusted for maximum light and the BF hole of the turret was set slightly off center to provide a bit of oblique illumination. The view, once I got it into focus (no small chore for me at this magnification), was a level beyond what I had experienced before. I saw obvious detail in the 7 diatoms with the largest striae intervals and if I squinted, and held my tongue the right way, I could even see the lines in Amphipleura pellucida. Of course, this detail wasn't present in the images but if you've been following this blog you know why. But I've gotta say, I'm pretty happy with the results after a bit of Elements work in removing the color and sharpening. The magnification of the following images is not constant.

Amphipleura pellucida  .27 microns... if you stare REAL hard you can see the lines

Frustulia rhomboides  .30 microns

Pleurosigma angulatum  .525 microns

Surirella gemma  .50 microns

Nitzschia sigma  .44 microns

Stauroneis phoenicentron  .72 microns

Navicula lyra  1.25 microns

Gyrosigma balticum  .67 microns

Bear hunting...

A tardigrade hanging on to some debris and checking out the surroundings

We STILL have two feet of snow over most of the ground up here in the Great White North and most of us would welcome even a hint of spring. Most of the few specimens I had in various containers were thinning out or getting boring. I needed some new specimens. Donning my Sorels, a warm jacket and a touque, I hiked into the woods near my place checking for moss in areas with some snow melt. Luck was with me and I cut off about 4 square inches of moss. Just for luck I also scraped off some light green lichen attached to some tree bark into a container. The temperature was -1C. Back home, samples of the algae and the lichen went into separate petri dishes and a small amount of distilled water was added to each. Impatience prodded me to keep checking for life but bed time arrived and I still hadn't found anything. Early this morning I squeezed some of the water trapped between algae fronds and checked it out under the microscope. Nothing.  Halfheartedly, I started looking through the lichen water and soon found my first tardigrade. Woo-hoo! Further searching found close to a dozen tardigrades, almost as many rotifers and one nemetode.

The above specimen appeared dead and when I tried to flatten him a bit to get more area in focus his skin broke and some of his inards spilled out. Check out the area near the back of his neck.

Frontal area of tardigrade

Here is a head shot that shows some of the details of the piercing and chewing apparatus. I've also uploaded a short video I took through one of my microscope eyepieces with a point and shoot camera.

http://youtu.be/1oHFeCgutdk

I also found a cuticle with 4 or 5 eggs in it but lost it while trying to transfer it to a slide. Must work on my lab skills.

Visiting with my Higher Power

I've got some of that high viscosity, RI 1.515 immersion oil, a 1.4 NA condenser, lens cleaning tissue, a Ph3 100X/1.3 Neofluar oil objective and the desire to experience something I haven't done before. In my excitement I forgot to set up Koehler illumination and in the end just moved the condenser and field iris around until I had something that resembles even illumination. I know you purists out there are just shuttering now but this is the way hobbyists often do things. I'll try to get it right next time.

First time out I just oiled the cover slip but couldn't find anything in the field. I went back to my 40X and sure enough, loads of micro-denizens but again my 100X couldn't find them. I regrouped, got out a new slide, put on a new drop of micro-critters and new cover slip and headed back. This time I centered a specimen with the 40 power objective first and then added oil to cover slip and condenser lens. When I rolled the 100X into place I saw a dark spot. After carefully fiddling with the fine focus I finally had an image and short lateral stroll down the slide delivered my first diatom. Wow! This view is definitely worth getting oil all over my microscope. Detail that was only hinted at before revealed itself with a clarity that was truly astounding. The only downside is the reduced depth of field; a price I'm willing to pay. I'll be reaching for that 100 power much more frequently now, especially once I fix up my camera issues. I'll attach a few photos and again apologize for the lack of crispness and focus but it is what it is for now.

The diatoms were about 50 microns long (I think) and viewed at 2000 power. I know, lots of empty magnification but I was after bling not doing research. After I settle down a bit I'll start doing things right again. All images were taken with phase contrast illumination.

Unknown diatom

Unknown diatom

Diatom frustule (Silica shell of dead diatom)

Unknown diatom

Filamentous Algae with bacteria (black spots)

Experimenting with Darkfield...

 My first attempts at Darkfield illumination (DF) a few weeks ago met with failure, in part due to misinformation on the internet (can that happen?) and lack of understanding on mine. I revisited the technique yesterday and much fell into place. Once I understood the stop had to obstruct ALL direct light going to the objective and the amount of direct light going into the objective is directly related to the NA. things became manageable.  There are methods available to calculate the exact size needed to obscure the direct light going into an objective but my stops were so easy to make that I just winged it. So let's get to work. I assembled a compass cutter, electricians tape, and a few old overheads from bygone presentations and a steel rule.

 The stops I was making are to be used with my 6.3X, 16X and 25X objectives  and I remembered reading about dimes and quarters being used so that became my starting point. Eventually, my 6.3X required something slightly smaller than a dime while the 25X needed a stop just larger than a quarter and the 16X was somewhere in the middle. 

The acetate circles were cut to fit my filter tray (32mm) and the electricians tape was added to just provide a black background when no slide was in place. Once you're finished just match the proper stop to it's companion objective, open up both the field  and aperture iris and be prepared to be amazed...especially, and perhaps only, if you have a suitable specimen on the stage. In my case I did, and I was totally blown away. I now understand what is meant when folks suggest the camera interpretation is nothing like the actual view. In my case, I saw what I might describe as diamonds thrown across a velvet sky were I the poetic sort. But I'm not so I'll just say the view was something out of this world. Perhaps a more suitable camera setup than mine can better represent what the eye really sees here but I doubt it. Here are a couple of images. Also you have to take into account the longer exposure required and the fact that these are motile diatoms...they move.

Pennate Diatoms in darkfield Illumination

DF image showing the large numbers of Daitoms in the FOV

 As the above image shows there were loads of these diatoms in my water sample, literally thousands within the area of the cover slip.

A few images...

I think the jury is now in on the status of my photo setup; a mixture of good and bad news. First the good...I'm able to record enough detail about what I see to recognize many of the features for future identification. The bad news is two fold. First the camera adapter is meant for a smaller, ccd video sensor and the projected image onto my sSLR sensor is quite small. Secondly, the adapter lacks the correction a normal Zeiss Kpl eyepiece makes to the objective image. All this means that at this point I'm not capable of taking good pictures but I can start recording my observations while I look for that *excellent* photo adapter. And as importantly, keep working on my photography skills.

Furthermore, today I discovered some open water on Sturgeon Creek and armed with a number of containers I scooped up several water, sediment and algae samples. An hour or so at the microscope coughed up a few specimens.

A Diatom Parade

Two pennate diatoms

Unknown

Unknown

Amoeba

An algae?

A cyanobacteria

Unknown

Unknown

A Filamentous Algae

A dead diatom

Vorticella (Peritrich ciliate)