Eppendorf 5315c micro-centrifuge

First trial of centrifuge on pond water 14/12/2017

“First Light” for my centrifuge – the second hand Eppendorf 5415c. I followed a procedure detailed on the Micscape website for using the centrifuge to concentrate the organisms in a pond water sample from Stowe Pool in Lichfield collected today.

The centrifuge worked well and did indeed concentrate the organisms (see photos below for procedure I followed) although the variety was limited in the sample.

Andy

1-The-Eppendorf-5145c-centrifuge

2-Pond-water-sample-including-piece-of-plant-root

3-Stirring-sample-to-release-organisms-into-water

4-Filtering-out-large-particles using tea strainer

5-Filling-Ependorf-1-5ml-tubes-with-pond-water – each one needs to have same amount fluid in order for centrifuge to balance:

6-Placing-Eppendorf-tubes-in-centrifuge- note that they are evenly spread around centrifuge to obtain balance at high G-forces – you should always use pairs and never one on its own:

7-Screwing-on-airtight-lid-onto-rotor:

8-Centrifuged-samples-on-homemade-stand:

9-Pipetting-off-excess-fluid-after-pellet-the idea is to carefully remove the fluid above the pellet of centrifuged solids at bottom of tube without disturbing the pellet. Ideally, the pellet and single drop fluid left after pipetting.

10-View-of-pellet-at-bottom-of-Eppendorf-tube. after pipetting – it is the brown spot at bottom of tube in picture. I used pipette to transfer the pellets and small amounts of fluid from 6 tubes to 2 tubes and centrifuged again to further concentrate the contents into two pellets:

11-Slide-after-pellet-and-drop-water-pipetted-onto-it-showing-concentrated-protozoa & debris. I pipetted the final 2 pellets onto a slide and the picture below shows many dark areas in the water drop – these are from the pellets:

12 Coverslip & nail varnish to seal it before viewing under microscope:

Photos of organism from sample today seen under Zeiss IM microscope x32 magnification. The first photo shows well one particular feature evident in many of the photos – which is that the centrifuge has caused chloroplasts to be concentrated on one side of the cell rather than spread throughout. Other photos show large vacuoles which I think are from damage to the cell. Most of the cells were inactive – I presume killed by the process, although the banana-shaped ciliated organism in the third photo below was alive and moving.

 

Eppendorf 5415 C Micro-Centrifuge Variable Speed with Timer – photos & manual

An old but sturdy piece of kit – useful for microscopy – allows samples to be concentrated (e.g. pond water samples) making it easier to find targets of interest. Only problem is that the timer does not work so I need to source one of those.

I have found a replacement timer – I intend to ask Ed whether he can take a look with me to see if we can work out how to replace it – not asked him yet – help please, Ed!
I also picked up a spare rotor – the screw on top was missing from mine.
All these problems with second hand stuff – its cheap but often needs work to get it working – but then that is the fun of the thing.
Below are photos of the centifuge, spare rotor and new timer.

Click below to read the manual for this machine (PDF file):

Centrifuge_Eppendorf_5415C manual

Andy

Diehl Series 600 T85 springwound timer – finding a replacement timer for the Eppendorf 5315c micro-centrifuge

Diehl Series 600 T85 springwound timer – finding a replacement for the 5315c centrifuge

Is this the correct one?

https://www.ebay.co.uk/itm/Poltic-or-Diehl-Springwound-Timer-30-MIN-Tanning-Bed-Type-600-T85-with-BELL/371656358069?epid=1376092377&hash=item568871f8b5:m:mjcQ-kmGpI3cfy5GJ-pL_oQ

or is this one a better choice?

https://www.ebay.co.uk/itm/Poltic-or-Diehl-Springwound-Timer-30-MN-Tanning-Bed-Type-600-HOLD/391491109081?_trkparms=aid%3D222007%26algo%3DSIM.MBE%26ao%3D2%26asc%3D49479%26meid%3Dac3b581f3858496789ad0cb084fb3974%26pid%3D100623%26rk%3D2%26rkt%3D6%26mehot%3Dag%26sd%3D371656358069&_trksid=p2047675.c100623.m-1

or this one?

https://spasandmore.com/catalog/timers/poltik-t85-typ-600-30-min-twist-timer-with-bell-8302.html

or possible UK source:

http://www.plccenter.co.uk/Buy/DIEHL/T85600?redirect=true