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#miCube: Modular fluorescence microscopy

miCube component overview

A full list of all components of the miCube is shown below. The numbers used in the tables are corresponding to the numbered items in Fig 1 below. The list is subdivided in main cube (red, nr 1-11), excitation path (blue, nr 12-18), and emission path (green, nr 19-30).

For all components, the design file (STL) is given if the component is home-made. All the files can be downloaded in a single file here (STL) or here (STEP). Full technical drawings of all components can be found in the supplementary information of our manuscript. If you want more detailed technical information, please mail us! For commercial components, the associated site is provided for more detail.

Fig 1: Overview of the individual components of the miCube. The individual components are numbered, and explained in detail in the tables. The colours correspond to the microscope category: red is the main cube, blue is the excitation path, and green is the emission path.

Main cube

Note that quoted prices are subject to change and are provided for indication only (Thorlabs being an exception here). For official qoutations, please contact the manufacturer or a distributor. Many parts such as cameras, optics, and lasers can be sourced from different manufacturers.
id Title Description Image Price
1 Glass plate insert
CNC milled - STL file
Size: 96-wells plate holder, 127.8 mm x 85.5 mm
€500
2 XYZ stage
An ASI XY stage (requires 4 mounting holes spaced by 75 x 125 mm) with a Optophase piezo Z-insert
€15.000
3 Objective €17.500
4 Top Cover
CNC milled - STL file
The Top Cover ensures correct height of the sample with respect to the parfocal distance of the objective (60 mm).
€500
5 miCube block The cube is milled out of solid aluminium to minimize drift. Practically all components connect to the cube in the end. €1.500
6 Neutral density filter
Thorlabs - NE60A-A
€65
7a Dichroic mirror holder
Thorlabs - DFM1/M
Magnetic hot-swappable dichroic mirror is placed inside.
€330
7b Dichroic mirror €500 to €550
8 TIRF filter €900 to €950
9 Connector for dichroic mirror and mirror
Thorlabs - C4W-CC
€45
10 45° elliptical mirror
Thorlabs - KCB1E/M and BBE1-E02
The fixed mirror ensures easy adjustment of position of the emission light on the camera by adjusting the two screws.
€270
11 Excitation path cover
3D printed - STL file
Ensures darkness of the setup.
-
Subtotal:
€37.110

Excitation path

id Title Description Image Price
12 Spacer
Thorlabs - SM1L05
€10
13 Reflective collimator holder
3D printed - STL file
Easy adjustment of height by tightening it to the mounting plate with two M6 screws.
-
14 Right-angle mounting plate
3D printed - STL file
-
15 Translation Stage
Thorlabs - PT1/M
25mm movement for aligning the excitation laser on the back-focal plane of the objective.
€260
16 TIRF lens
Thorlabs - AC254-200-A-ML
TIRF lens to focus excitation light on back-focal plane of objective. 200mm focal length.
€90
17 Aperture
Thorlabs - SM1D12SZ
€70
18 Reflective collimator
Thorlabs - RC12APC-P01
A parabolic mirror in the collimator ensures a well-collimated beam simply by attaching the fibre to the collimator.
€880
- Laserbox
Omicron - Lighthub 6
Custom laser combiner containing 4 lasers at 405 nm (60 mW), 488 nm (200 mW), 561 nm (500 mW), and 642 nm (2x 200 mW).
€55.000
Subtotal:
€56.310

Emission path

id Title Description Image Price
19 Camera €15.000
20 Camera mount
3D printed - STL file
Camera mount specifically designed for the Andor Zyla 4.2 Plus sCMOS, along with the used tube lens in this setup.
-
21 Astigmatism block
3D printed - STL file
An astigmatism holder containing an astigmatism lens can be placed on two positions.
-
22 Astigmatism lens
Thorlabs - LJ1516RM or LJ1144RM
Astigmatism lens with 1000 or 500 cm focal distance for optional super-resolution astigmatism.
€110
23 Astigmatism lens holder
3D printed - STL file
A single lens can be slot into place with a screw. A magnet can optionally be added at the bottom for a more solid placement in the block.
-
24 Astigmatism lens holder
3D printed - STL file
A single lens can be slot into place with a screw. A magnet can optionally be added at the bottom for a more solid placement in the block.
-
25 Emission path cover
Thorlabs - SC600, cut to length
Ensures darkness of the microscope.
€30
26 Connector for tube and filter block
3D printed - STL file
-
27 Emission filter €325
28 Emission filter holder
3D printed - STL file
A single lens can be slot into place with a screw. A magnet can optionally be added at the bottom for a more solid placement in the holder.
-
29 Tube lens holder
3D printed - STL file
-
30 Tube lens
Thorlabs - ITL200
200 mm focal length
€425
Subtotal:
€15.890
Total:
€109.310

Laser triggering

The triggering of the lasers is performed with a home-made Labview software SMILE (see image below), in combination with the hardware listed below. For now, further information or access to the software is available upon request. This hardware can be used for stroboscopic alternation laser excitation schemes (sALEX) as demonstrated in Farooq & Hohlbein, Phys. Chem. Chem. Phys., 2015.

Id Title Description Image Price
- DAQ
National Instruments - PCIe-6353
X Series Multifunction DAQ (32 AI, 48 DIO, 4AO), 1.25 MS/s single-channel sampling rate (781049-01)
€3.000 +
- Connector Block (2x)
National Instruments - SCB-68A
Noise Rejecting, Shielded I/O Connector block (782536-01)
€1.000
- Cables
National Instruments - SH68F-68F-EPM Cable
€350

Accessories

id Title Description Image Price
- Filter holder
3D printed - STL file
Can hold up to 10 emission filters or astigmatism lenses. Has two holes at the bottom for easy placement with M6 screws.
-
- XYZ cover with LED light A single IKEA INREDA LED spot with the connector cut off is soldered to a 1/8 inch DC jack. The DC jack is connected to a circuit that controls the voltage of the spot using a potentiometer and a transistor. The schematic is depicted below (click to enlarge): L1, L2 and L3 resemble the IKEA LED array, L4 is an indicator light for the power switch. A 16V DC switch-mode power supply (SMPS) was used. As the INREDA spot normally runs on 12V DC, R2 is placed in series with the potentiometer to limit the maximum voltage coming from the transistor Q1. The LED array is connected via J2 to J1 on the Brightfield Control Box. -
- Top Cover alternative
CNC milled - STL file
Alternative for the normal Top Cover (nr 4). Does only hold the Z-piezo, without the XY stage.
€500
- Breadboard
Thorlabs - MB3060/M
Aluminum breadboard, 300 x 600 mm. All components (except the laser box) are placed on this breadboard.
€240

3D printing settings
id Printer Material Support Adhesion Layer height (µm) Top/Bottom Thickness (mm) Wall Thickness (mm) Infill %
13 / 14 UM2+ PLA Yes Brim (5) 200 1.0 1.2 20
26 UM2+ ABS Yes Brim (5) 200 0.8 0.8 20
21 UM2+ PLA Yes Brim (5) 200 1.0 1.6 20
23 / 24 / 28 UM2+ PLA Yes Brim (5) 200 1.0 1.6 20
20 UM2+ PLA Yes Brim (5) 200 0.8 0.8 20