Archive for the ‘Cell Culture’ Category

How to build a ghetto micro-environmental chamber for inverted fluorescence microscopy.

September 8, 2011

This project is very easy, super cheap, and has a very specific application.

Background

Our lab has been trying to figure out ways to cheaply set-up an environmental chamber for time lapse imaging of tissue cultures.  Environmental chambers are typically expensive or just a pain to make and so I threw this ghetto chamber together.  This chamber works for us because we use an inverted confocal microscope.  This make-shift chamber is opaque and will not work for any set-up where light passes through the bottom of a sample and out the top (in our case light passes up through the bottom and then is scattered back down).

The chamber requires an appropriate gas supply (our goal is to use air with 5% CO2) and some way to regulate the flow to the chamber.  We are having trouble locating a good gas supply so the chamber hasn’t been tested out yet.

 

What you will need

    • Equipment
      • drill
      • drill bit
    • Reagents and consumables
      • a pipette box lid that is larger than culture plates
      • barbed fitting with threaded end
      • tubing (runs from fitting to tank)
      • washer
      • nut

 

Required consumables.

Below is what the inside of the pipette box lid looks like.  This is also the orientation the lid should be in when it is drilled.

Pipette tip box lid (bottom view)

Drill a pilot hole (use a smaller bit than your final size and if needed increase from one size bit to the next until the final bore size is reached).  The pilot hole is important because pipette tip box lids are likely to crack when being drilled.

Pipette tip box lid with drilled hole

Now the chamber can be assembled.  Push the threaded end of the barbed fitting into the chamber.  Place a washer and then a nut onto the portion of the threaded barb that is on the inside of the chamber.  The figures below shows the finished chamber on the outside and inside.

Micro-environment chamber assembled.

Inside of the micro-environment chamber.

Naturally a longer tube than the one I have depicted above will need to be used to reach the air tank.  The short piece I have attached is there for display.

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3D model – 24 well plate for cell culture

August 12, 2011

You are probably thinking, “Well hey, this here post has nothing to do with cheapass science!”  and you would be almost right.  I made the 3D model for a 24 well cell culture plate because I wanted to use it as a guide in my design of something that is cheapass science related.

Awhile back I blogged about a crude suction device I fashioned for allowing me to drain 24 well plates more quickly (four wells at a time versus one by one).  Since then (and even before then) I had been dreaming about using a 3D printer to make the object (along with other fun things).  But the reality is that I do not have a spare $1500.  But this money issue won’t hold me back because I recently found out that it is rather inexpensive to get small objects 3D printed in plastic!

To go about making the tool I decided it would be best to create a 3D representation of a plate.  I did this first because it should make it easier for me to plan and visualize the tool I am drawing.  Once I have the tool designed it should be a mere trifling to have a company prototype it for me.

If you are interested in downloading the 3D model for the 24 well plate, go here.

Cell Spreader

April 15, 2011

Can’t find your cell spreader?  Too cheap to buy one?  Did someone indefinitely borrow yours?

Well, if you have access to a Bunsen burner and 9″ pasteur pipettes you can quickly make a spreader of your own.

Before:

After:

Additional examples:

Video:

The video below shows how to warp the pipette with heat into the shape of a cell spreader.

More information:

  • First pass; place the pasteur pipette into the flame and after a few seconds the tip of the pipette will bend downwards via gravity.  This is because the heat weakens the glass and the portion of the pipette which is in the flame will act like a pivot point.
  • The pipette will very quickly cool and you can near immediately put it back into the flame.
  • Second pass; as before, put the pipette into the flame and it will bend downwards via gravity.
  • Third pass; place the very tip of the pipette into the flame and melt it shut.  If it is not closed off, the tip will fill up when you sterilize it with alcohol and will spark.