Open Blink Max

Overview

Max is a open-source dual-color TIRF microscope with a high-power laser combiner. Why is it called Max? Because the first microscope at KG lab has the nickname of Willy/prism, a reference to the Dutch monarch family.

The laser box is composed of four lines, three of which are multimode diode lasers (405 nm, 488 nm, 638 nm), and one DPSS laser (561 nm). To increase the power at 638 nm, we employed two lasers that are combined with a polarisation beam splitter (PBS). The laser beams are delivered through a multimode optical fiber agitated to reduce the speckles. A motorised motor facilitates the switch between epi, HILO and TIRF. For the main frame, we adapted the Micube to hold our sample stage and high NA objective.

We use micro-manager 1.4 gamma for the image acquisition and control of most hardwares except the lasers. The lasers are modulated via an Arduino and Digital-analog converter (DAC), and controlled by customised script.

Max is inspired greatly by K2 TIRF at Ganzinger lab, the LaserEngine at Ries lab, and this flat-field illumination implementation. We thank the Open Life Science program that helped us with the documentation of this project.

You could find the CAD model of the system (laser box and the main body), as well as the laser control (wiring and scriptis) at TBD

Schematic

Laser Box

The laser box is contained within a 450*450 mm breadboard (MB4545/M) with enclosure. All lasers are raised up to 70(?) mm height with customised aluminum heat sink.

Model numbers are for Thorlabs unless specified otherwise.

id	Title	Description	Image	Supplier, Model	Cost in €
1	Diode lasers 405 nm	500 mW, with driver	Diode laser and driver	Lasertack, LAB-405-500	687
2	Diode lasers 488 nm	2000 mW, with driver		Lasertack, LAB-488-1000	3310
3	Diode lasers 638 nm, 2x. One is oriented perpendicular to the other	2x1000 mW, with drivers		Lasertack, LAB-638-1000	866
4	DPSS lasers 561 nm	1000 mW, with analog modulation(0-5V)		CNI laser, MGL-FN-561	11150
5	Dichroics	1" longpass dichroic mirros to combine the laser lines		Thorlabs, DMLP425/505/550/605	708
6	PBS	1" polarisation beam spliter, mounted by KM100PM/M and PM4/M, in order to combine two 638 nm lasers	PBS	Thorlabs, PBS251	315
7	Fiber launcher	A 1/2" 19mm ach. doublet (AC127-019-A) to focus the combined beam into the fiber. Mounted in 1" tubale lens tube (SM05V10, SM1A6, SM1M15, SM1TC), FC/PC adaptor(SM1FC)	Fiber launcher	Thorlabs	220
8	Multimode fiber	70*70 um square profiled MM optical fiber, NA=0.22		Ceram Optec, Customized	350
9	Vibrational motor	9000 RPM vibrational motors that are taped onto the optical fiber to break down the speckles in the output beam profile	Vibrational motor Vibrational motor mounting	Hackerstore.nl	4

Laser box control

Since all lasers in the laser box take 0-5V analog signal as instensity modulation signal, we used DAC chips and Arduino UNO to control the laser lines. You could found the scripts run on Arduino IDE, where simple interactive control is realized by serial communication between the PC and Arduino UNO. The wiring between the Arduino, DACs, and the laser drivers is a prototype on breadboard, and has been running in KG lab. Interlock and laser safety measures should be implemented in any circumstance.

Excitation

id	Title	Description	Image	Supplier, Model	Cost in €
1	Fiber adaptor	Couple out the beam at the output of the optical fiber mounted on cage plate (CP33/M)		Thorlabs, SM1FC	30
2	Laser collimator	A 30 mm achromatic doublet to collimate the beam at the output of the optical fiber. Mounted in 1" cage after the fiber adaptor	Laser collimator	Thorlabs, AC254-030-A	85
3	Laser cleanup filter	Filter out the autofluorescence from the MMF		Chroma, ZET 405/488/561/640xv2	575
4	TIRF stage	1D motorised stage scanning the laser beam across the back focal plane(BFP) of the objective	TIRF stage assembly	Thorlabs, KMTS25E/M	1500
5	TIRF lens	150 mm achromatic doublets to focus the beam onto the BFP of the objective, mounted on CXY1A		Edmund Optics, 147-643	283
6	Elliptical mirror	Mounted on KCB1E/M and assembled with the TIRF lens on the TIRF stage		Thorlabs, BBE1-E02	70

Microscope Body

id	Title	Description	Image	Supplier, Model	Cost in €
1	Modified MiCube	Adapted from MiCube. Reduced in size to accommodate 150 mm focusing lens	Micube	in-house CNC milled	1700
2	XYZ stage	Nano precision sample stage		Smaract, CLS5252	14000
3	Objective	Oil immersion objective, NA = 1,5. Mounted on the micube with M27 to RMS adaptor (RMSA3)		Olympus, UPLAPO60XOHR	12400
4	Quad-band dichroic	Dichroic mirror under the objective, mounted on DFM1/M and fixed on the micube with screws on the back		Chroma, zt405/488/561/640rpc	575
5	Elliptical mirror	Mounted under the dichroic mirror with KCB1E/M and C4W-CC	QUad-band dichroic and mirror mounted inside of Micube	THorlabs, BBE1-E02	65
6	Sample holder	Home-made slide holder, mounted onto the stage		In-house CNC milled	\

Detection

We adapt the detection scheme in K2 TIRF at Ganzinger lab for our dual channel detection.

id	Title	Description	Image	Supplier, Model	Cost in €
1	Tube lens	180 mm tube lens compatible with Olympus objectives		Thorlabs, TTL180-A	686
2	Notch filter	Filter out the excitation laser beam		Chroma, ZET405/488/561/640m-TRFv2	1000
3	Slit	Semi rectangular slit adjustable from two directions, cropping the image to fit in a single channel on camera. Alternative could be Owis SP60	Slit	Recycled from MadCityLab RM21
4	4f lenses	Three 300 mm achromatic doublet mounted on LMR1/M	Two channle 4F system	Qioptiq, G32 2336 322	300
5	Dichroic mirror	Long-pass dichroic splits image into red and green channels, glued on compact KMS/M with picodent Twinsil		Chroma, ZT640rdc-UF3	450
6	Square mirror	Reflects the green channel with maxium surface, mounted on compact KMS/M		Thorlabs, BBSQ1-E02	74
7	Emission filters	Band-pass filters in the red and green channels respectively		AHF, ET685/70, ET595/50	650
8	Camera	sCMOS camera		Teledyne Prime BSI Express	10800