Photographing Chromosomes

As readers of my blog will be aware, I enjoy imaging natural things. For me ” Natural World” includes the very big (like galaxies) and the really rather small (like cells). I’ve decided to include some Microscopy (or Photomicrography) in my blog, so by way of a start I thought I’d include some images of chromosomes, the data bookshelves of life on our planet.

Chromosomes require fairly substantial magnification and the simplest way to achieve acceptable resolution at x1000 with a standard optical microscope is to use oil immersion microscopy; this is how the following images were taken …

 

Chromosomes during Mitosis

Mitosis is the process of duplicating & splitting the nucleus / genetic material of a eukaryotic cell. It is normally followed by cytokinesis which completes cell division, resulting in 2 identical daughter cells. Mitosis is used for growth & asexual reproduction, it is distinct from meiosis (used for sexual reproduction) and binary fission of prokaryotic cells. Other than the ‘normal’ interphase state of cells, Mitosis has 4 main phases & these allows us to view & photograph chromosomes:

  • Prophase – During interphase the genetic material will have been duplicated, now the nucleus swells and the chromatin within, coils up to form chromosomes; each consisting of two chromatids. Simultaneously the centrosome duplicates & each resulting diplosome moves towards opposite ends of the nucleus.
  • Metaphase – In most cases the nucleus has now become indistinct. The chromosomes line up across the centre of the cell and protein spindle fibres develop across the cell, coordinated by the centrosome at either side of the cell.
  • Anaphase – Each chromatid pair splits, forming sister chromosomes. The protein spindle threads now pull the chromosome sisters apart, to opposite ends of the cell.
  • Telophase – Each group of chromosomes are now at either end of the cell, they are now enveloped by a new nuclear envelope. The chromosomes now uncoil back in to chromatin and are no longer distinct for us to see.

To complete cell division the cells cytoplasm will now cleave and two new sister cells will form; this is cytokinesis.

So what of it? Well the chromosomes are particularly distinct during Metaphase & Anaphase and that gives us a great opportunity to photograph them. The process doesn’t take very long, Metaphase can be over in 15 minutes; so we need to look at a whole bunch of cells & pick out the few that have been ‘caught in the act’ as it were. The micrograph below is of onion cells dividing mitotically near the root tip …

Mitosis - Plant Click image for a larger view …

The image was taken at 1000 times magnification (x10 ocular, x100 objective) and used oil immersion to improve the image (see below). The cell in the centre of the picture displays distinct blue stained chromosomes and just possible to make out is the occasional spindle fibre. Two other cells can be seen at different stages.

 

Giant Chromosomes in Fruit Flies

Drosophilia, the favourite of geneticists across the ages are one of a few insects that have an unusual characteristic. They develop ‘giant’ or polytene chromosomes in their salivary glands. DNA is replicated many times, without cell division taking place. This can lead to large banded chromosomes with perhaps 1000 times the normal compliment of DNA, 85% of which may be concentrated in to the dark bands. These give us another excellent opportunity to photograph chromosomes.

Drosophilia Polytene Chromosome  Click image for a larger view …

Image details: Magnification x1000, Light Microscopy with Oil Immersion, Canon 300D  ISO 100  TV 1.3s, 4 image stack using CombineZM

 

Technique & Equipment

How were these images taken? The basic equipment is pictured below:

_MG_8488 Click image for a larger view …

Referring to the image above, from bottom to top: Light Source, Slide, Oil, Objective Lens, Ocular lens, photo adapter, DSLR

ie. Immersion Oil is carefully placed here (picture below), between objective lens & slide coverslip.

_MG_8484 Click image for a larger view …

The microscope is carefully focused away from the slide until best focus is found. One draw back of this method is that the focal plane is absolutely tiny. The advantages are simple, at high magnifications light microscopes loose resolution due to refraction at the glass air boundaries; by using immersion oil with the same refractive index as the glass, 2 such boundaries are removed & resolution is restored.

I shall add more microscopy images in time to come.

 

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2 comments on “Photographing Chromosomes

  1. Shannon Scott

    Wow, that looks like a lot of fun. I’ve thought about getting into home photo-microscopy, but don’t know where to start (at least without spending loads of money). Could you suggest a microscope (I can’t identify the make/model of the scope you’re using)? Are there any resources (websites or books) that you found useful when starting out?

    Thanks,

    Shannon.

  2. antamuk

    Hi Shannon,

    Yes its lots of fun and unlike astrophotography, I don’t have to wait for the clouds to clear! You don’t need to spend a fortune either; my microscope is made by Brunel Microscopes, its a Wedmore SP14. You’ll find its details at the bottom of the following page: http://www.brunelmicroscopes.co.uk/student-microscope.html

    Brunel also sell 2nd hand scopes; if you’re not in the UK, I expect there’ll be a similar manufacturer in your part of the world too. As for starting off help, Brunel sponsor the Microscopy UK group. Their web site contains lots of useful info, just follow the link below:
    http://www.microscopy-uk.org.uk/index.html

    You’ll need to think about what camera to use, there are adapters available for both DSLRs & compact digital cameras, so you should be able to get going with whatever you already have.

    Finally you can either purchase pre-mounted specimen slides or make your own – I do a bit of both.

    I hope this helps, have fun.
    AnnMarie

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