Leica DMI 4000 tutorial

Transcription

1 Leica DMI 4000 tutorial Before using the Leica DMI 4000, You will need to put down your name on the reservation system = 1

2 Welcome to the Leica DMI4000 Microscope tutorial How to start up the system (p.3) How to use the stand DMI4000 (p.5) How to use the stand from the soft (p.10) The Differential Interference Contrast (DIC) (p.11) Fluorescence (p.12) How to acquire an image and its different adjustments (p.13) How to focus easier (p.17) How to acquire several images and overlay them (p.18) How to see, to save your images and their information (p.21) How to annotate your image (p. 22) How to shut down the system (p.23) 2

3 System Start up (step1) Turn on the Fluorescence lamp (Xcite) Turn on the Microscope button You have to turn on the light only if it is cold (approximately 30 minutes after its last use) 3

4 System Start up (step2) Turn on the PC Introduce your Username Introduce your password Start the Leica Application suite software 4

5 Microscope ( left side) Intensity of halogen or fluorescence lamp (+/-) (INT) Condenser Aperture for the contrast (+/-) (AP) Fluorescence to transmitted light selection 5

6 Microscope (front part) Eyes piece position Camera position Shutter for the fluorescence (Xcite) Filters (BLUE/GREEN/RED/DEEP RED) 6

7 Microscope (right side) Choose your objective manually: HCX PL S-APO 5x 0.15 HCX PL APO 10x 0.30 N PLAN 10x 0.25 HC PL APO 20x 0.7 IMM CORR λbl HCX PL APO 63x OIL To put a magnifying lens (1.6 x) To move the stage 7

8 Microscope (up part) Field diaphragm, for Köhler adjustment (close/open) condenser 8

9 Check your sample through the eyes piece Select the objective For bright field On the left side, Press on the TL/BF button Setup the intensity, with the buttons INT setup the aperture diaphragm, with the button AP Focus the sample For fluorescence On the front part, Select the filter (BLUE / GREEN / RED / DEEP RED ) Press the shutter Focus the sample 9

10 Microscope on the soft Under Acquire and Mic1, you can see the objective that you re using You can change the kind of contrast in transmitted light, or select fluorescence For Fluorescence, you can change filters and press the shutter You can select the eye piece or camera position For Acquiring your image after all settings done 10

11 Differential interference contrast (DIC) Under Acquire and MIC1, select a DIC contrast You can change the aperture diaphragm and the intensity of the light To change and choose the right contrast, turn this button 11

12 Fluorescence Under Acquire and MIC1, select FLUO, so you can setup the field diaphragm Setup the intensity of the fluorescent light Change the filters Press the shutter 12

13 Acquire an image Under Acquire and Camera, you can adjust the exposure time The contrast (gamma) 13

14 Histogram and saturation options Under Acquire and Camera and Histogram, You can see the grey scale You can tick Show Over/Under Exposure, and you will see the over exposure in red and the under exposure in blue, you shouldn t have some red dots on your picture You can also reduce the grey scale 14

15 The Input options Under Acquire and camera and input option, you can choose if you want some pseudo color The bit depth The right format (HQ 1600x1200) 15

16 The binning Under Acquire and Camera and Input option, You can choose a binning, it will increase the camera sensitivity, reduce the exposure time But you will lose some resolution 16

17 How to focus easier Under Acquire and Camera and Region of interest, If you select zoom focus, it will zoom the region you want so you can easily find the right focus 17

18 How to acquire several images and overlay them Under Select Acquisition Mode, select Image Overlay The window lambda will be opened With the play button, you can start and stop the live and the shutter 18

19 To do the overlay and save the configuration For each channel you have to select the filter Choose the exposure time Choose a pseudo-color You can save your configuration When it s done you can Acquire your overlay 19

20 To see the overlay image Under Browse and λ, you can see Each channel And the overlay picture You can also tick the images that you want to see in overlay mode and click on overlay show 20

21 How to see, to save your images and their information Under Browse and file information, you can see the information linked to the image Under Browse and File directory, you can see in which file your images are saved 21

22 How to annotate your image Under Process and Annotate, you can choose to note some information about the image The name, description, date and time The scale bar Lines, measurements (ex. distance line) To save the picture with the information, you have to select Merge. Be careful, if you do it, you can t go back. So if you need the original picture, copy it before you annotate it 22

23 How to shut down the system If you was using oil/glycerol objective, clean the objective with ethanol/water Before shutting down the system, you have to look if there is somebody using this microscope after you. In this case you haven t to shut it down but you have to log off your session Shut down the PC Turn off the microscope Turn off the light 23