MiMic

Photolitography Lab

Ex1
We want to produce in vitro an array of functional cardiac muscle myofibers using
micropatterning techniques [1]. We need to fabricate a PDMS mold to perform µCP on soft
substrates. The pattern of proteins that we want to create to culture cells is an array of
rectangular lanes with the following features:
Protein
The PDMS mold should have features 100 µm
L= 1mm tall. You must use a NEGATIVE PHOTORESIST
L
w= 75µm (see SU8 µ-series DataSheet). The Mask Aligner
d= 100µm to expose the resist is equipped with a 1000 W
mercury arc lamp that has an emittance of 6,5
d w mW/cm2 (measured).

• Draw the mask (transparent/black parts)

• Decide the parameters to produce the wafer
10x 40x
C2C12 cells after 2 days of C2C12 immunofluorescence
culture reached confluence analysis to detect desmin (red)
confined inside the laminin and cellular nuclei with
lanes Hoechst
1.Cimetta, E., et al., Production of arrays of cardiac and skeletal muscle myofibers by micropatterning techniques on a soft substrate. Biomed(blue).
Microdevices, 2009. 11(2): p. 389-400.

R. VISONE
Exe 1
5 lines h = 100 µM
Ꙍlamp = 1000 W
Lamp irradiance = 6.5 mW/cm2
NEGATIVE PHOTORESIST

• Draw the mask (transparent/black parts)

y • Decide the parameters to produce the wafer
1 2 3 4 5
x

• Draw the mask

Lateral view mask

1 2 3 4 5

z
PDMS
z
1 2 3 4 5 su8
x
x
wafer
PDMS

Top view
Surface mask

x
• Protocol to produce wafer

1) COATING
h= 100um → decide which SU8 → SU8-µ50

Spinning curves Speed (rpm) Accel (rpm/sec) Time (sec)

1st ramp 500 100 10

2nd ramp 1000 300 30

2) Sb process → thickness SU8 = 100µm

65°C → 95°C → 65°C
5 min → 20 min → 5 min

3) EXPOSURE → dose → exposure time → check the thickness

From the table → 240 mJ/cm2

Time = exposure energy / irradiance = 240 (mJ/cm2) / 6.5 mW/cm2 = 36.9 sec

4) Peb → thickness = 100µm

65°C → 95°C → 65°C
5 min → 10 min → 5 min
5) DEVELOPING → Thickness = 100µm

Development time = 10 min

 MiMic
Photolithography Lab

Ex2
Design a PDMS chip to study neuronal growth in response to chemical factors [2]. We need to
provide a culture chamber to host the cellular bodies (soma compartment), some µchannels
for guiding axonal growthing (axonal µgrooves) and a compartment filled with conditioned
medium (chemical factor compartment). You must use a NEGATIVE PHOTORESIST (see SU8 µ-
series DataSheet). The mercury lamp has an emittance of 8 mW/cm2.
Top View SU8
Soma chamber: µgrooves: Factor chamber:
Axons L= 5mm h2= 50µm L= 600µm L= 5mm
Factors

Som w= 500µm w= 5µm w= 100µm

a Lateral View SU8 d= 20µm h2= 50µm
Y z h1= 10µm
Soma Axons Fact
x µgrooves x

• Draw the mask (transparent/black parts)

and decide the printing resolution (DPI)
• Write the protocol to produce the wafer 40x
Microfluidic-based culture
with all the parameters platform to direct axonal
growth of CNS neurons Axonal growth is guided through the
microgrooves
2,Taylor, A.M., et al., A microfluidic culture platform for CNS axonal injury, regeneration and transport. Nat Methods, 2005. 2(8): p. 599-605

R.
VISONE
Exe 2 Soma chamber: µgrooves:
L= 5mm h2= 50µm L= 600µm
Axons w= 500µm w= 5µm
d= 20µm

Factors
h1= 10µm
Soma
Lateral View SU8 Factor chamber:
Y z L= 5mm
Soma Axons Fact
w= 100µm
x µgrooves x h2= 50µm

Ꙍlamp = 1000 W
Lamp irradiance = 8 mW/cm2

• Draw the mask (transparent/black parts) and decide the printing resolution (DPI)
• Write the protocol to produce the wafer with all the parameters

• MASK & DPI

1) MASK for Soma + factors 2) MASK for µGrooves

mask mask

SU8 SU8

wafer wafer

Mask 1 Mask 2

Resolution → smallest features that we have 5µm = µgroove

10x resolution → 1 dot = 0.5 um

DPI = 1 DOT / 0.5 um = 2 dot/um = 2/ 3.9 10-5 inch = 51282 DPI 1 inch = 2.54 cm
• Protocol to produce wafer
2 step process → start with the smaller h → start with µGrooves
• µGrooves h= 10µm → SU8µ25
1) Spinning curves Speed (rpm) Accel (rpm/sec) Time (sec)
1st ramp 500 100 10

2nd ramp 4500 300 30

2) Sb process → thickness SU8 = 10µm

65°C → 95°C → 65°C
1 min → 5 min → 1 min

3) EXPOSURE → dose → exposure time → check the thickness

From the table → 130 mJ/cm2
Time = exposure energy / irradiance = 130 (mJ/cm2) / 8 mW/cm2 = 16.25 sec
4) Peb → thickness =10µm
65°C → 95°C → 65°C
1 min → 5 min → 1 min

5) DEVELOPING → Thickness = 10µm

Development time = 4 min

• Soma & factor chamber h= 50µm → SU8 u 100

1) Spinning curves Speed (rpm) Accel (rpm/sec) Time (sec)

1st ramp 500 100 10

2nd ramp 4000 300 30

2) Sb process → thickness SU8 = 50um
65°C → 95°C → 65°C
2 min → 6 min → 2 min

3) EXPOSURE → dose → exposure time → check the thickness

From the table → 160 mJ/cm2
Time = exposure energy / irradiance = 160 (mJ/cm2) / 8 mW/cm2 = 20 sec

4) Peb → thickness =50µm

65°C → 95°C → 65°C
1 min → 6 min → 1 min

5) DEVELOPING → Thickness = 50µm

Development time = 5 min

Recommended Program:
- Spin at 500rpm for 10 sec with an acceleration of 100 rpm/sec (1st ramp)
- Spin at desired ω for 30 sec with an acceleration of 300 rpm/sec (2nd ramp)