Microscopy

Materials

Light microscope with camera & appropriate software

Slide(s) with samples

Protocol

Turn on power source, laser (may not be necessary for bright field, only for fluorescence), and microscope.

Open camera software and ensure cameras are on.

Place the slide upside down on the stage and secure between stage clips (do not flip left-to-right) — images seen through the microscope will be inverted.

Ensure the camera pin is pushed all the way in (allowing light to transmit to the eyepiece) and turn on the light.

Select desired magnification level (10x, 20x, etc.) and use the joystick to move the slide in the x-y plane so the correct portion of the sample is under the objective.

Look through the eyepiece and focus using the large knob (large vertical stage movements) then the small knob (small vertical stage movements), being careful not to bring the objective and slide too close.

Pull the camera pin all the way out (allowing light to transmit to the camera).

Select the appropriate camera (black and white or color) and press live camera.

Ensure the sample remains focused through the computer screen and adjust exposure so cells are visible and the background is bright.

Click “snap” to take a picture, and save to the appropriate folder on the computer.

File name should include the experiment name, slide number, eye number, magnification level, and date.

When finished, close the software, return the microscope to the lowest-magnification objective, push the camera pin all the way in, and turn off the light and microscope, then the laser, then the power source.

Results

Focused, visible photos of the samples in .czi format. Transfer via flash drive; convert .czi to .jpg with Fiji or similar to open on other computers.