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How to Mount Zebrafish in Methylcellulose

Using Methylcellulose is convenient for mounting live embryos for observation under the compound microscope.  Methylcellulose is a plant extract that is dissolved in fish water to make a viscous solution.  It puts a relatively loose hold on the embryo, so you can move the embryo to different positions.  Because of this, embryos that are old enough to move MUST be anesthetized for photography.  It is typically used with glass depression slides and NO cover slip.  It is very gentle and non-toxic, and the embryos can easily be recovered; thus, it is typically used for LIVE embryos. 

Chemical safety:  No hazards are listed for methylcellulose, and no protective gear is required, but you may want to wear gloves.  If it comes in contact with your skin or eyes, wash the affected area with soap and copious amounts of water and inform the instructor.

1. Using an embryo loop or a small wooden stick [cold, 4 degree Celsius], place a small dab of 3% methylcellulose (see recipe below) in the center of the depression of a glass depression slide.  Don't just drop the methylcellulose on, push it down, otherwise it will float away.  If it does float away, then just push it back into the depression of the glass depression slide.


2. Dechorionate and/or anesthetize the embryos as necessary as listed in “How to use anesthetics on zebrafish.

3. Place the embryo on top of the methylcellulose, and layer fish water on top to fill the depression.


4. Move the slide under the stereomicroscope and gently push the embryo into the methylcellulose with an embryo loop until it is secure.  Use the loop to position the embryo in the orientation that you want. Be careful to keep the embryo covered with water during your observations or it will dehydrate and die.


Recipe for Methylcellulose

1. For 3% methylcellulose it is equivalent to 3g methylcellulose in 100mls of fish water.
2. Rock overnight in a rocker until the solution is dissolved.
3. Split the solution into 50 ml tubes and freeze the solution until needed.


4. When the solution is needed, thaw for usage (use heat for time purposes to help methylcellulose go into solution).