Leica DFC500 Kurzanleitung

Warren Lab JMW, v3 06/2013

OPERATING NOTES FOR THE LEICA MICROSCOPES AND CAMERA

Shut-down computer when not in use. This may improve long-term camera performance.

Camera

•DFC500 with 12MP 2/3" CCD chip and Bayer color filter.

•The camera can be used with either microscope.

•The camera connects to the computer by a Fire wire cable and has no power switch. The

camera LED light indicates the status as follows:

oLED is red when PC Powers up or camera is first connected.

oLED turns green, then flickering orange as LAS opens, as it opens to the Acquire tab.

oLED turns green in all LAS tabs apart from Acquire, where it is flickering orange.

oLED goes green and back to orange a few times as you Acquire an image.

oLED remains green when LAS is closed.

•To transfer the camera between the two microscopes:

1. File > Exit LAS (do not quit the entire program)

2. Configuration > Hardware Configuration; select correct configuration.

3. File > Exit (fully quit program; this will turn the camera off)

4. Loosen the set-screw that holds the camera in place.

5. Carefully move the camera over to the other microscope, making sure the Firewire cable

does not get caught on the monitor.

6. Tighten the set-screw to thumb tightness.

7. Place dust cap on the empty port of the other microscope.

Camera Trouble-shooting

•If saving photos at high resolution, do not put too many photos in a folder, as the camera will

take a long time to save.

•The LAS will crash if photos have too long an exposure, due to issues with the DFC 500

camera. In general, take exposures <800 ms.

Stereomicroscope

•Leica M165C with 0.63x and 2x objectives

•Base plate has several options: polarized, glass, or black/white for picking. These plates are

exchanged by pressing on the lower rim to lift the plate out.

•The entire square black plate can be removed and replaced with a large glass plate. This is

done by loosening the back screw with a coin and pressing on the back right corner to lift the

plate out.

•Use care when moving the plates. If working with rock samples, use and index card between

the sample and the plate to protect it from scratches.

Warren Lab JMW, v3 06/2013

Petrographic microscope

•Leica DM2500 with polarized transmitted and reflected light

•Objectives: 2.5x, 5x, 10x, 20x, and 50x

•Beneath the stage are the polarizer, daylight filter, aperture diaphragm, and condenser.

•Use the condenser for 10x mag and above; it provides diffusive illumination known as

Kohler illumination. The height of the condenser is adjustable, but should not need to be

moved. To properly position the condenser:

1. Focus on sample with analyzer in or out and using an intermediate objective (10x or 20x).

The position of the condenser should be approximately the same for all objectives. Close

the field diaphragm down until it falls just within the field of view. Adjust the height of

the condenser until the diagram edges are in sharp focus. (The field diaphragm always

shows the conjugate image planes, so the edges can be visible, whereas the aperture

diaphragm shows the image plane and cannot be seen.)

2. Check the centering using the 2 set screws located by the aperture diaphragm. One screw

moves the position NW-SE and the other moves NE-SW. The field diaphragm should

again be closed down so that the ring is just inside. Move the set-screws until the aperture

diaphragm is centered.

•Smith reflector - part of the reflected light path that directs light down and maintains the

polarization of the light. The turret containing the Smith reflector has 4 positions but the

others are empty and the turret should always be left in position 4.

•The condenser path for reflected light is aligned and focused in a similar manner to

transmitted light. In this case, the field diaphragm in the reflected light path is aligned using 2

set screws. Centering of this is similar to centering the transmitted light using the field

diaphragm. However, in reflected light there is no focusing that is done.

•The aperture affects the contrast of the image seen in the microscope. The aperture

diaphragm in transmitted light has color-coded markings that indicate the optimal setting for

each of the objectives. It should be closed down at higher magnification. The aperture

diaphragm in reflected light can also improve image contrast.

Warren Lab JMW, v3 06/2013

Camera Software – Detailed Notes

•The software is referred to as the Leica Application Suite (LAS).

•Right-click when starting the software, to run in “Administrator” mode, which gives more

options and runs the camera faster than “User” mode.

•After moving the camera, you must tell the software which microscope you are using. To

switch between hardware configurations, first exit the LAS by going to: File > Exit LAS.

Then select the correct microscope by going to Option > Select Hardware Configuration.

Then quit the LAS software and re-open.

•The software has various tabs:

oSetup - you should never need to modify this!

oAcquire ("Live" mode) - this will always display a live image when the software is open.

The tab for "Mic" detects the microscope, the zoom and the iris. Loading the different

hardware configurations will modify the contents of this tab. The software can auto-

detect the zoom and objective for the stereomicroscope. On the petrographic microscope,

the objectives are not auto-detected but there are pre-set scales for the different

magnifications. The tab for "Camera" contains all other settings for taking

photomicrographs and is described in detail below.

oBrowse - Post-image processing.

•File names - Can either enter each time or select auto numbering:

Options>preferences>image tab> (un)check always confirm image tag

•Save images as Tiffs as these are completely uncompressed.

Specific settings to check

•Always do a white balance after moving the camera

•Stereomicroscope: make sure the boxes for horizontal and vertical flip are checked under

processing

•Petrographic scope: make sure the boxes for horizontal flip are checked under processing

•Petrographic scope: Check the box "Confirm Microscope Setting" to always check the

magnification/objective when taking a photo.

The various options under "Camera":

Auto Exposure – useful for getting roughly in the right range, but in general you will always need

to adjust this manually. You should aim to have the brightest areas be just at saturation (white

but not burnt out). This can be adjusted either with the slider or by using the arrow keys on the

Warren Lab JMW, v3 06/2013

keyboard. When auto exposure is turned on, adjusting the exposure is done by adjusting the %

brightness.

Exposure – when auto exposure is turned off, then the actual exposure (in ms) can be changed.

The roll wheel on the mouse can be used to adjust the exposure.

Gain – you will probably never use this. It is a way to speed up the camera frame rate, but this

occurs at the expense of the image resolution. The default setting is 1.

Color Saturation – this adjusts the color intensities. With a good white balance, there should be

no need to adjust this. The default setting is 1.

Gamma – a form of contrast. Lower values correspond to less contrast. The default is 0.6.

White Balance – controls the color of "white" which depends on the illumination source. The two

microscopes have very similar white balances, but you will always need to adjust this after

moving the camera. The stereomicroscope has LED transmitted light and LCD light source for

the gooseneck illuminators. Both of these give a relatively blue light. The petrographic

microscopes uses halogen bulbs, but both the transmitted and reflected light have daylight filters

to make the light bluer. To do a white balance, use a white sheet of paper and in manual mode

draw a box around the white area. For transmitted light, this can instead be done using a glass

slide or by adjusting the white balance in reflected light mode. (The ideal material for a white

balance is 70% grey, but a white card works well.)

Show Camera Settings – this provides access to an older version of the LAS interface. You

should have no desire to use this.

Show Over/Under Exposure – this will highlight areas of the image that are over-exposed (red)

or under-exposed (blue). Click on an under- or over-exposed area to assess the balance and use

the mouse scroll wheel to adjust the exposure. Also have the histogram window open to assess

the exposure balance.

Processing – shading imbalance can be adjusted for using the shading correction feature. This

can adjust for variations in the lighting; which is very useful on the stereomicroscope as the

goosenecks can provide uneven lighting. In transmitted light on the stereomicroscope, the

camera optics go through the right eyepiece and pick up slight shading from the base plate on the

left side of the image. To create a shading correction, use a flat empty field such as a defocussed

sheet of paper or the glass (polarized) base plate in transmitted light. Create a shading reference

and give it a name (e.g., "Left Edge Correction"). Once this is created, you can toggle between

the correction mode and the correction free mode.

Resolution of Line Format – this should be left at full resolution. Sometimes during teaching it

can be useful to increase if the refresh rate is too low. The default setting is 1360x1024, 1 shot

interpolated, which will read every pixel on the chip.

Warren Lab JMW, v3 06/2013

Capture Format – Moving color filter (Bayer mosaic filter) relative to the CCD when taking a

picture, which requires 2 green, 1 red and 1 blue images to be taken. Therefore, the filter must

move 4 times to get all 4 images. The default setting is 1360x1024, 4 shot co-site color. A larger

number of shots will result in better resolution images but the file size will be huge.

Input Options – A particular camera setting can be saved as a default setting:

Input options > configuration > create new default setting

A variety of pre-defined contrast settings are available, but these are not of much use. The option

exists to turn off the chip cooling, but do not do this. The chip will perform better if cooled,

though it does not have to be cooled when not in use. When the software is not open, the cooling

fan is turned off.

Histogram – Show the signal display, which can be manually adjusted. However this results in

loss of information as reduces each color element from full 255 spectrum. You should not have a

reason to adjust the histogram.

Region of Interest – Can crop by drawing a box and only acquiring the image within the box.

Can zoom focus to magnify an area to aid in focusing.

Check Color – this can be adjusted after doing a white balance.

Calibration Settings – Calculated indicates that the scale-bar is chosen based on the microscope

specifications. For the stereomicroscope, this is automatically detected. For the petrographic

microscope, this must be manually selected to match the objective.

HDR Imaging – HDR stands for high dynamic range. This takes multiple exposure images to

capture brightest and darkest segments of the image. Heath (JH Technologies) is supposed to

provide more information on this, as it is a recent upgrade.

Left Click on Image – allows you to draw a box and brings up menu options. Can use to select a

white balance area or for zoom focusing.

Right side buttons:

Pan window – allows scrolling around when zoomed in; Zoom in/out; Fit to screen; 1:1 - restores

image to full resolution.

Scale-bar – Can adjust parameters, location. Acquiring an image does not necessarily save the

image unless the box "Merge" is checked. This can be done in post-processing by selecting

Merge, which will either replace the image or save a second file.

Dual Image Browser – Can either compare live image to saved image or compare two acquired

images.

Warren Lab JMW, v3 06/2013

Image Mosaic – This allows an image mosaic to be collected. The software can automatically

detect the location of overlaps. It works best with 20-30% overlap. Use the dual viewer when

taking images to monitor the overlap between images.

Form – all image data (metadata) is displayed here. This information is saved as a separate text

file with the image.

Record Details – Full microscope data associated with image (extended version of form data)

File Formats

Saving a photo with scale bar and other annotations creates variable numbers of files in addition

to the tiff:

CAL.XML extension is the Calibration and Microscope Information

ANX extension is the Basic Annotation Data

EAX extension is the Extended Annotation Data

LMD extension is the Interactive Measurements Settings and Data

SBX extension is the Scale Bar Settings Data

SNR extension is the Store and Recall Data

THB extension is a Thumbnail Image

Windows Shortcuts

•Ctrl+Shift+Esc brings up the Task Manager

•Windows+e brings up the file explorer

To Do (6/2012)

•Currently running LAS 4.2.

•New camera driver for DFC500 may be released soon (according to Phoebe Tsellos,

11/2012). This might address some of the issues with the i5 CPU being recognized by the

LAS software installed on these types of computers (alternatively, need a firmware update

for the computer).

•If camera problems persist, consider getting a new FireWire cable, as currently have an A

(camera) to B (computer) connection. A connection of A to A would work better, though also

depends on the metal alloy used in the cable. Should check with current Leica representative

(Shawn Hussey) before doing this.

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