Weak fluorescence, insufficient brightness | Leica DM IRB specification

Possible errors

Weak fluorescence, insufficient brightness:

Wrongly stored, overaged or faded specimens.

Fast fading of the specimens (e. g. for FITC).

Unspecified filter combination.

Numerical aperture of the objective too low.

Eyepiece magnification too high.

Spent lamp.

Room too bright.

Trinocular tube: wrong beamsplitter setting.

Stray light due to reflections at the condenser.

Low-contrast image due to:

Excitation bandwidth too wide.

Inspecific staining.

Fluorescing mounting medium.

Autofluorescence of objective or immersion oil.

Glass surfaces dirty.

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Image 87

Leica DM IRB manual Weak fluorescence, insufficient brightness, Glass surfaces dirty

Contents

Leica DM IRB Tel. +49 Leica DM IRB Copyrights Contents Automatic lowering Operation Text symbols and their meaning Important notes on this manual→ p Mains plug must only be inserted into a grounded outlet General safety informationPage Intended application Key subassemblies Microscope and its components Tube StandFirst of all, here is a general overview Stands Tube mount Coarse and fine control Brightness adjustmentMains switch Condenser height adjustment IR/R tube adapter Specimen stages and accessoriesLamphousings Objective nosepiece and objectives Ambient conditions Temperature 10 36 C Relative humidity Installation site Unpacking Installation Assembly Assembly toolsAssembly of the transmitted light illumination column Transmitted light illumination column, back view Assembly of condensers Condenser 0.30 S70 Position condenser disc, empty Condenser 0.90 S1 bottom up Assembly of IC condenser prisms Condenser top Assembly of condenser 0.30 S70 Condensers 0.53 S23 and 0.90 S1 Assembly of condenser holder Assembly of filters and filter holder Assembly of field diaphragm Retrofitting individual IC prisms Examples of prismsPrism a for objectives N Plan 5x Differences between prism D and D1 Inserting the analyserInserting the polariser Fluorescence module Inserting the fluorescence module Mirror housing and illumination telescope Connect the lamp plug to the connecting socket in the stand Assembling and exchanging incident light lamps Lamphousing 107 L Lamphousing 106 z L Lamphousing 106 z L Type Power units It is extremely important to heed the follow- ing advice Assembly of the tubes and tube adapter IR/R + PR = with and without back reflection Bino HC BSA 25 Trino HC FSA 25 P and PR Tube adapter R/IR HC, 2 Clamp screw Adapter, 6 Photo/TV port Transformer Edge of the mirror housing, 11 Mirror housing Inserting the eyepieces Inserting graticulesScrew the retainer ring back Inserting the photoeyepieces Screwing objectives in and outRemove the screw covers from the objective threads Only press the spring at the side Plate x/y stage Rotary stage and insert frame for coverslips Connecting the microscope to the mains E version DM IRB/E Features of the Leica DM IRB/E Important noteFunction and operation Controls Assembly and initial installation Learn mode Installing the objective prismsExit option in the display flashes I T Installing the objectives Display Phase contrast Display Objective magnificationDisplay IC objective prism Operating modes Dry/Immersion Immersion objectivesTo learn further objectives Oil immersion objectives Exiting the Learn modeParfocality Dry objectives Sw i t c he s Individual user adjustments Installing the fluorescence filter cube Concluding the installationWith the key, → b Motorized objective nosepiece Now appears in the LC display at the bottom right Changing the operating modeOperating modes DRY and Immersion Automatic lowering of the objective nosepiece Brightness adjustment Controls of the electronic focus are Electronic focus Latter flashes if the combination is wrong Coded IC objective prisms option Motorised fluorescence filter cube change option Control unit Connecting the control unit Component Name on the unit Keys on the control unit Lamp Focus Examples for the use of the footswitches Person sensor option Switch position AutoSwitch position Light on Switch position Sensor OFF Front controls 86mm S 10 0 xPH3 HH Switching on the halogen lamp Operation Basic setting Focusing the specimen Adjustment specimen Operation of L objectives with correction mount Setting the tubes and eyepieces Only when one eyepiece is without an adjustable eyelens Side of the tube Trinocular tube HCI 3T22 Operation of the front photo/TV port Operation of the side photo/TV port Locking objectives Operation of objectivesImmersion objectives Colour coding of objectivesPage Fig.Ê 62Ê Ê Setting Koehler illumination Replace the eyepiece or disengage the Bertrand lens Possible errors Phase contrast observation Operation of phase contrast Setting phase contrast with condensers 0.53 S23 and 0.90 S1 Darkfield observation Operation of transmitted light darkfield Crossing the polarisers DL polarisation TL interference contrast Operation Transmitted light interference contrast Objectives for ICT Centration of the condenser prisms Setting ICT contrast Sources of error if ICT image quality is unsatisfactoryChoice of prisms Fluorescence observation Operation of incident light fluorescence Alternative centration methods Method Centration with a centring aidMethod Centration in the rear focal plane of the objective Lamphousing 106 z L with halogen lamp, Xe and Hg lamps MethodCentration of 12 V 100 W halogen lamp Hg 100 W and Xe 75 W lamps Hg 50 W mercury lamp Halogen lamp Hg 50 lamp Xe 75 lamp Centring the field diaphragm Centring the aperture diaphragm Glass surfaces dirty Weak fluorescence, insufficient brightness Light filters Operation of filters Scale 10 mm = 100 divisions Operation of the slide overlay deviceSlide overlay device Slides with the following line patterns are available Fig.Ê 73ÊMacroÊ device on FSA 25 PE tube with tube adapter Operation of the macro device Field of view Ø 10x/20 eyepiece Total magnification in the eyepiece would therefore be Calibration Length measurementsFollowing components are required for length measurements Transmitted light stage micrometer for cali- bration With zoom magnification Vario TV adapter MTV = objective magnification x tube factor Monitor diameter Fig.Ê 80ÊLeicaÊ DM IRB, equipped with three TV cameras Long-term video microscopy Operation of LMC Principle Principle of LMC S40/0.50 LMC condenser ComponentsComponents LMC consists of the following components Screw the LMC objectives into the objective nosepiece Assembly/adjustmentAssembly AdjustmentPage Areas of application Care and maintenance Cleaning objectives Acids, alkaline solutionsRemoval of immersion oil On/off switch does not respond no illumination TroubleshootingMechanical errors Electric errors 106 Additional fluorescence lamp does not respond 107 846-205.000-00Name Schutter FST 108 Ê Lamphousing 106 z, opened 109 Replacing the 12 V 100 W halogen lamp in lamphousing 106 z Changing the Hg and Xe lamps on lamphousing 106 z 111 Transport Storage 0Ê -Ê Technical descriptionPerformance data of objectives Objective lettering 114 Or A, B, CObjective type performance class Plan 10x/0.22 Colour coding of the objectives White WaterLocking of objectives Performance data of eyepieces Decisive value is always the smallestEyepiece field of view Eyepiece graticules Field performance of objectives CB1.5, CB3 Filter performance data Use Grey/neutral density filter NGreen filter, GR, panchromatic BG20 Like HCI B22, but with variable viewing angle of 15Ê -Ê Tube performance dataBinocular tube HCI B22 Binocular ergotube HCI BV22 Tube adapter R/IR Trinocular tube HCI 3T22 121 Condenser performance data Condenser 0.30 S70 Condenser disc Object guide122 Stages Plane stage and mountable object guide Lamphousing performance dataLamphousing 106 z Lamphousing 107/2 C., stabilised/non-stabilised TypeHg ultra high pressure lamp 50 W A.C Xe high pressure lamp 75 W D.C., stabilised Overvoltage category Contamination class 125 General technical dataGeneral technical data Fuses Ambient temperature 10Ê -Ê 36C Relative humidity Technical data of the power unit General technical data Technical specificationsULa = lamp voltage 127 Main wearing and spare parts, tools Product name EU Conformity declarationPage Ernst-Leitz-Straße