Pond, lake, stream and river water (fresh water) microscopy

Second attempt at oblique dark field illumination of algae from neighbour’s pond using halogen swan-neck illuminator on Zeiss Standard microscope

This time I used my centrifuge to concentrate the sample – snow outside/cold means number algae per ml in the water low.

Bright field images of algae x40 objective – unfortunately they show that I have some work to do aligning the optics as lot of colour fringes…there is also quite a lot of dust on the optics of this microscope – I need to give it a good clean! However, not all out of focus rings are dust – much of it is algae in different planes on this live sample.

I particularly like the second and third pictures as they show long cilia from the spherical organism.

Video from this session showing motile organisms:

Dark field using oblique illumination – set up with Zeiss Standard microscope:

I have found that the best dark field is when the fibre-optic tips are placed on the stage pointing virtually horizontally at the end of the objective.

Dark field with x10 objective, using above equipment – looks like a star field in the telescope! Can you recognise the constellations?

The above two images came directly from the camera.

The following are the same two pictures but this time I have used curves in GIMP to remove part of curve below the data and hence blacken the background:

Dark field using x40 objective – this is where I am breaking new ground with success at dark field using x40 objective. So far, using my Zeiss IM microscope, I have been able to obtain excellent dark field images using the 10x non-phase objective and a phase annulus, but the higher power objectives don’t seem to work so well using that system. I think maybe the NA on the objectives is too high compared to the NA on the condenser, but am not sure of the reason.

I found that if I varied the position of the swan neck heads to direct the light more downwards (angled the lights to point down rather than horizontally) then this varied the lighting effect. The lighting is no longer true dark field but is still interesting! Example picture below:




Commercial Vorticella slide

Vorticella on a commercial stained slide, viewed using Zeiss IM microscope at different magnifications. Damian and I have previously seen Vorticella live in local pond water samples – see previous posts.

I used the Bresser MikOkular camera in “new” trinocular head (second hand from ebay) – this differs from previous trinocular head in that this is the one that is recommended in the Zeiss IM microscope handbook. I noted that the previous head, although it works, has small black ring around outside of field of view that I assume means field stop is too small for scope. This new one does not have this. This new one also provides 23mm ocular attachment on trinocular port, into which the Diagnostic Instruments adapter fits directly without needing a clamp.

I also tried out a dark field condenser on microscope today – did not work well – not sure why – so photos below are back to the phase condenser that came with the microscope, used without phase annulus (i.e. in bright field mode).


x10 objective:

x32 objective:

x63 objective (slide upside down so light only has to go through coverslip in this inverted microscope – 63x objective has only limited working distance). This is a panorama of 17 panes, joined using Microsoft’s Image Composite Editor:

Damian’s views in his Lomo Biolam microscope

Damian used his renovated Lomo Biolam microscope to view the small organisms we found in our recent trip to Branston water park and cheek cells we collected from his mouth and stained with H&E staining on 2/2/2018.

Andy and Damian

Cheek cells x20 objective

Cheek cells x40 objective

X90 objective. All these images taken afocally from one side of the binoviewer on the Lomo, using hand-held Samsung S7 phone through x10 ocular and objective mentioned. There is also 1.5x multiplication effect from the binoviewer.

Therefore the magnification on image below with x90 objective = 10 x 90 x 1.5 = 1350 x. Contrast and brightness tweaked in the image editor on the Samsung S7 phone – no other processing:

The following photos are all with x10 objective, otherwise as above. They are of organisms from Branston water park:

The following picture shows Damian taking pictures through the ocular using the a ocal imaging technique of holding his camera up to the ocular.

Microscopy sample taken from edge of lake at Branston Water Park 20/2/2018 viewed 22/2/2018

Damian collected this sample 20/2/2018 when Andy & Damian went there. In the previous post, we described how we centrifuged this sample to get it ready for microscopy. In this post, we show microscopic images from the slides we produced.

  • Bresser Mikrocam 5.0 camera.
  • Zeiss IM microscope.

Andy & Rhys & Damian

Photo of large ciliated organism x20 objective bright field:

Photos x32 objective – mixture bright field and Phase Contrast images showing bacteria, diatoms, and something I don’t know!

Videos – Phase Contrast x32 objective:

Microscopy open water trawl net samples from Branston Water Park collected 20/2/2018 viewed 22/2/2018

The following images are from the sample that Damian collected 20/2/2018 on our walk to Branston Water Park viewed through the Zeiss IM microscope.

I had kept the sample fed with sugar dissolved in water over last 2 days and in sunlight and exposed to oxygen (opened top bottle).

All images with Bresser Mikrocam 5.0 5MP camera.

The black circles are bubbles.

Specimens killed with 10% formalin (20ml poured into 2 litre bottle) and then filtered with coffee filter to concentrate the specimen. Specimen mounted using water-based mountant.

I am particularly proud of the first photo. Darkfield achieved using Ph2 Phase annulus on Zeiss IM microscope with x10 objective and then I used GIMP to darken background a little (I used curves).


Darkfield image x10 objective Phase 2 annulus:

Same field as above, bright field, x10 objective:

Following images are all brightfield, x2.7 objective:


Microscopy of stream sample collected 18/2/2018 from stream between Netherstowe School in Lichfield and adjacent allottments viewed on 22/2/2018

The sample I took from the stream between Netherstowe School in Lichfield and adjacent allottments on 18/2/2018 has been kept in a jar and fed with a spoonful of sugar dissolved in warm water each day (to keep bacteria growing – these are bottom of food chain for larger predatory protozoa). I keep top open to allow oxygen in. The jars were kept next to our french doors to allow light to shine in each day.

Today I used phase contrast on the Zeiss IM microscope to view these images – so images and video below are either in bright field (without phase contrast) or Phase contrast using annulus Ph1 on the Zeiss. All images and video are using x32 long working distance objective and taken with Bresser 5MP Mikrocam camera.


Phase contrast images, x32 objective – show bacteria and a protozoal organism:

Bright field (first) and Phase contrast (Ph1) image (second) of same field to compare:

Video of sample x32 objective – this is brightfield not phase contrast:



Flat worms & things: Microscopy sample of water from Branston Water Park, Lichfield Road (A38), Burton Upon Trent DE14 3HD

We went to Branston Water Park for walk and collected two samples:

(I) Trawl sample collected using homemade plankton net

(II) Sample from around water plants near edge + some plant parts to include surface-living protozoa.

Andy & Damian

Damian uses my homemade Plankton net to collect sample from Branston Water Park lake (I used a filter for an air conditioning system for the net – to give the required pore size which is measured in microns! Those protozoa are very small):

Creating centrifuged pellet from sample from around water plants next to edge lake:

Video of live microscopy of centrigue pellet from Branston Park lake water sample using x32 objective:

I am very proud of this video – definitely one worth watching!! Look out for the two layers of cilia on the one organism and the vortex-producing mouthparts in another clip, sucking in small organisms as food like a Dyson vacuum cleaner!

This first video is a compilation of highlights from the second video that follows afterwards:

Full video – shows a greater range of organisms and behaviour than the highlights video above –  if you are having difficulty accessing the embedded video below the URL for direct link to the video on YouTube is https://youtu.be/SKpROQCVp0A)


Photos from live microscopy of centrifuge pellet x32 objective:

Two flatworms:


I am having some difficulty identifying this. I can get it down to three types of genera but can’t be more specific. Note the two layers of cilia (hair-like projections):

The following looks like algae but is motile – the video shows two of them moving like liners on the sea:

I am not quite sure whether this organism is the same as the ciliated organism above or not. This one is free swimming whereas the other was (I think) attached at one end to some debris:

Is this following the same as above? This one sucked in debris and small organisms like a Dyson vacuum cleaner as I watched. It is on the video but was more impressive at the eyepiece, as the little doomed organisms could more easily be seem being sucked at high speed into the mouthparts (right side of organism in middle of cilia below):


Brightfield microscopy of stream sample next to Netherstowe School, Lichfield collected 18/2/2018 observed 20/2/2018 using Zeiss IM microscope

I prepared a brightfield live sample (unstained) of the water I collected from the stream between Netherstowe School in Lichfield and the allotments next door to the school on 18/2/2018 today 20/2/2018 and viewed this using my Zeiss IM microscope (transmitted light).

Bresser Mikrocam 5.0 camera


All following photos & video using x32 long working distance objective – some protozoa and bacteria visible: