The Lumicks C-Trap is a correlative force-fluorescence microscopy system that allows simultaneous manipulation and visualization of dynamic single-molecule interaction. A unique combination of multi-trap optical tweezers, multi-color, and label-free imaging modalities, and a microfluidic device enables high throughput spatiotemporal manipulation, measurement, and visualization of diverse biophysical processes.

The C-tap has a powerful (1064 nm, 20 W) trapping laser and uses a high NA objective lens (WI NA 1.3 and OI 1.49) to generate up to four simultaneous, continuous-wave optical traps. The stiffness and position of each optical trap can be controlled independently to provide maximum flexibility to the user such that a wide range of single-molecule experiments can be performed. The trap can be manipulated with an extremely high position accuracy (<0.2 nm step size) that ensures the user can perform force-distance, constant distance, and constant force measurements with a Brownian-noise limited accuracy.

The C-Trap is equipped with dedicated fluorescence microscopy capabilities enabling a correlated, precise, and stable sample manipulation, force measurement, and single-fluorophore detection. TIRF microscopy offers single molecule fluorescence imaging at the surface with a high background rejection resulting in a high fluorescent signal-to-noise ratio. The platform also includes a bright-field microscope for simultaneous accurate live bead-tracking.

The C-Trap includes a microfluidics solution that facilitates repeatability in single-molecule experiments in a laminar flow environment. The microfluidic flow cell provides multiple adjacent laminar flow channels that do not mix. Samples are connected to a specific channel in the flow cell and can be independently switched on and off through automated fluidic valves. The lack of actual physical barriers between adjacent channels enables users to swiftly move the trapped objects across the channels, form in-situ assembly of single molecules, and carry out experiments under different buffer conditions.

FEATURES:

1. Stable and precise sample manipulation

Traps: 4 independently steerable traps with higher force resolution (<0.1 pN @ 100 Hz) and stability (<0.3 pN over 2 minutes) facilitates flexible molecular configurations. The trap can be operated as a force clamp with a maximum escape force of >1000 pN and is equipped with temperature control module (room temperature to 45  ͦC).

2. High throughput experimental setup

Both the surface and solution-based experiments streamlined by a high-resolution piezo-controlled nano stage and a 5-channel µ-Flux laminar microfluidics system. Custom-prepared chambers on a glass slide can also be used.

3. Diverse and correlated imaging capabilities

TIRF: Offers single-molecule fluorescence imaging (488 nm, 561 nm, and 638 nm laser lines) with a high background rejection. CMOS camera detector (Peak QE 95%) with the complete area of interest captured simultaneously, providing a fast video rate imaging (> 25 Hz) solution for fluorescent single molecule surface assays.

Widefield (HILO): Switchable between TIRF and Hilo (dirty TIRF) modes to facilitate both single molecule surface assays as well as dumbbell constructs in the sample volume.

IRM (Interference Reflection Microscopy): Label-free 3D imaging allows image acquisition of nanoscopic objects on the surface at a higher rate (25 fps @ 10 frames avg).

4. User-friendly control and fully correlated data acquisition

Bluelake: Compact, integrated control software that facilitates a fully correlated data visualization and acquisition.

Lakeview: Simplified and readily available data visualization and analysis software platform (download Lakeview program). The users could also export the data and perform the visualization/analysis using Pylake, a Python-based package.