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Using U-Shaped Glass Capillaries for Sample Mounting

Prepare specimens for light sheet imaging

Light sheet microscopy tilescan of zebrafish. DLS_Sample_Preparation_zebrafish_tilescan_intro.jpg

The DLS microscope system from Leica Microsystems is an innovative concept which integrates the light sheet microscopy technology into the confocal platform. Due to its unique optical architecture, samples can be mounted on standard glass bottom petri dishes and require little or no adaptation when compared to conventional mounting procedures. Here, we present a convenient way to prepare specimens quickly for light sheet imaging.

Introduction

In light sheet microscopy, specimen preparation is a crucial topic. Ideally, it should be as simple and fast as possible. The DLS system has the advantage that it allows for horizontal mounting and easy sample acces. Specimen preparation has only two requirements: the sample needs to be placed in the focal plane of the detection objective (between the TwinFlect mirrors) and slightly elevated from the petri dish bottom, so that the mirrors/light sheet can reach the distal part of the sample. 

Figure 1: Position of the sample in the free area sample space of the detection objective.

U-shaped glass capillaries, which are customized for different TwinFlect mirror sizes, help users fulfill these specimen preparation requirements in an intuitive manner.

Optimum positioning of the sample by using u-shaped glass capillaries.
Figure 2: Optimum positioning of the sample by using u-shaped glass capillaries.

Workflow

The video clip will lead you through the sample preparation workflow using u-shaped glass capillaries

Materials and methods

U-shaped capillaries are available in two different sizes:

  • Outer diameter = 1.5mm; inner diameter = 1.03mm, length 20mm (Order number 158007061, 50 capillaries) → fit TwinFlect 2.5mm, TwinFlect 5mm and TwinFlect 7.8mm
  • Outer diameter = 2.5mm; inner diameter = 2.04mm, length 20mm (Order number 158007060, 50 capillaries) → fit TwinFlect 5mm and TwinFlect 7.8mm

With a length of 20mm, the u-shaped glass capillaries are ideal for use with 35mm glass bottom petri dishes (e.g. Greiner, ibidi, Matek,…).

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