Super-resolution Microscopes

PlexFlo Multiplexing Platform

Accessory for sequential labelling in localization microscopy applications

PlexFlo Multiplexing Platform

Applications are developing in localization microscopy where the number of targets for imaging far outstrips the number of probes that can be chromatically separated. Sequential labelling with microfluidics allows a much larger number of targets to be addressed. Bruker microfluidics accessory units are designed to allow sequential labelling to be incorporated into localization microscopy protocols with the Vutara VXL single-molecule localization microscope.

 

Bruker offers two units—PlexFlo and PlexFlo96. The units are controlled by Vutara's SRX software and allow flexible configuration of fluidics sequences that are run in conjunction with imaging cycles to allow users to label an unlimited number of probes and image them in a sequential fashion.

Sequential Labelling with Ease

PlexFlo

PlexFlo contains 15 buffer reservoirs that can be swapped out during an experimental workflow, enabling the multiplexing of many labels in one experiment. These distinct features make PlexFlo optimal for DNA-PAINT applications.

PlexFlo96

PlexFlo96 enables even further multiplexing capabilities, as it contains 96 buffer reservoirs that can be swapped during one experiment. Consequently, PlexFlo96 is ideal for applications that require a large number of labels in one experiment, such as chromatin tracing.

Informatics

The probe associated with each sequence has a specific color that is user-defined, and that is used as a color scale for visualization of experiments. During analysis with SRX, individual probes can be analyzed, or analysis can be carried out that compares probes.

9-step OligoSTORM walk spanning 8.16 Mbs along both homologs of human Chromosome 19. Courtesy of Guy Nir and Ting Wu, Harvard University

Applications

The Bruker microfluidics unit can be used with Vutara to address any localization microscopy application where a large number of targets needs to be imaged. Fifteen probe vials are provided, and the system allows probe vials to be interchanged during experiments, thus allowing an unlimited number of targets.

Example localization microscopy applications:

The Bruker microfluidics unit is also compatible with the Opterra confocal for serial labelling confocal applications.

Specifications

  PlexFlo PlexFlo96
Samples/Reservoirs (small volume) 15 x 2 ml vials (Eppendorf standard) 96 well in microwell plate
Wash/Buffer Reservoirs (large volume) 1 x 100 ml, 1 x 50 ml, 2 x 15 ml 8 x 125 ml
Waste Container 1 x 100 ml GL45 bottle  
Temperature Control of Sample No 39○ - 99○F; 4○ - 37○C (microwell plate)
Sample Swap During Experiment Yes Yes
Pressure Driven Flow Control Yes Yes, and vacuum for extraction from 96 well plate
Pressure Range 0 – 345 mbar (±0.1% CV above 10% full scale, ± 105 µbar resolution)  
Flow Sensor Thermal based, 1 µl/min to 1 ml/min (±1% CV)  
Software Integration Yes Yes
Automatic Cleaning Procedure Yes Yes
Dimensions 370 mm x 300 mm x 204 mm  

Software

Control of the unit is via SRX software. The software has a flexible interface module (Fluidics Manager) for the set-up of fluidics. An unlimited number of sequences can be set up, each with its own unique steps.

 

The Bruker microfluidics accessory is a unit designed to allow sequential labelling to be incorporated into localization microscopy protocols with the Vutara VXL single-molecule localization microscope. The unit is controlled by Vutara's SRX software and allows flexible configuration of fluidics sequences that are run in conjunction with imaging cycles to allow users to label an unlimited number of probes and image them in a sequential fashion.

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