Snouty High NA Single Objective Light Sheet

FP BASE

Microscope Built By Tanner Fadero

Optics

Objectives

• For Viewing
• CFI Plan Apo Lambda S 60XC Sil 1.3 NA 0.3WD
• FOV: 400 um diameter XY, 20 um Z
• pixel size at sample: ~0.102 um
• Lightpath/image forming
• AMS-AGY v2.0 NA1.0 Lamda 450 -700nm  ∞/0 WD=0
• CFI Plan Apochromat Lambda D 40X 0.95NA (with coverslip glued to lens)

Intensity Filter Wheel

1. ND Filter OD 3.0 (0.1% of the light through)
2. ND Filter OD 2.0 (1.0% of the light through)
3. ND Filter OD 1.5 (3.16% of the light through)
4. ND Filter OD 1.0 (10% of the light through)
5. ND Filter OD 0.5 (31.6% of the light through)
6. ND Filter OD 0.0  (100% of the light through)

Dichroic

1. zt405/488/561/640rpcv2-UF1 (Chroma)

Emission Wheel

1. ZET405/488/561/640mv2  (Chroma)
2. ZET488/561m (Chroma)
3. ET445/58m (Chroma)
4. ET535/70m (Chroma)
5. ET630/75m (Chroma)
6. ET706/95m (Chroma)
7. ET750sp-2p8 (Chroma)

Lasers

• VersaLase (8) system For Light Sheet
• 405nm, 100mW
• 488nm, 200mW
• 561nm, 50mW
• 639nm, 200mW
• Triplet Depletion Laser
• Stradus 808nm, 250mW

Transmission light path

• Brightfield
• Differential phase contrast

Hardware

• Oko Stage Top Incubator with CO2
• Adapters for 96 Well plate, standard slide (25mm x 75mm), 35mm Dish, and Lab-Tek II Chambered Coverslips
• Hamamatsu Orca-Quest 2 Camera
• Prior Pure Focus system
• NanoScan SP400 Piezo Sample Scanner with 400 micron travel range (in process of being set up)
• Galvo mirror for light homoginizer
• Lightpath/image forming
• AMS-AGY v2.0 NA1.0 Lamda 450 -700nm  ∞/0 WD=0
• CFI Plan Apochromat Lambda D 40X 0.95NA (with coverslip glued to lens)
• OptoSpin filter wheel 6 positions 32mm filters (emission) from Cairn
• Six-Position Motorized Filter Wheel for Ø1” (Ø25 mm) Optics (ND filters for lasers) from Thorlabs

PC

• Asus Pro WS WRX90E-SAGE SE
• AMD Ryzen ThreadRipper Pro 7965WX sTR5 24C/48T 4.2GHz 128MB 350W
• 384 GB RAM(DDR5)
• 1 TB NVMe M.2 SSD.

Software

• MicroManager 2.0
• how to cite Micro Manager: https://micro-manager.org/Citing_Micro-Manager

### Imaging Reccomendations

• See slide and dish recommendations here: Plates and dishes for imaging, Note Must be No. 1.5 Coverslip equivialant
• Imaging dish footprints that can be used:
  • 96 well/plate foot print
  • Standard slide (25mm x 75 mm)- include coverslip bottom well slides.
  • 35mm coverslip bottom dish
  • 60mm coverslip bottom dish

Data Management

• Expect lots of Data!
• To help manage your data efficiently:
  1. Back Up Locally
  • Immediately back up your data to a personal hard drive (USB 3.0 recommended).
  • Please do not leave data transferring on the Snouty when other users are scheduled.
  • Once your backup is complete, immediately remove your data from the computer.

  OR

1. Upload to the CALM file server
   • The Snouty is directly connected to ~petabyte server. Instructions to access the server can be found here.
   • The server offers a 3-month grace period, this is not permanent storage.
   • You can access the server on the UCSF network or via VPN.
   • The server is only available to internal users.
   • Pro Tip: The CALM offers analysis workstations with 10 Gb/s high-speed access to the mic.ucsf.edu file server. These can be booked through iLabs for data transfer, viewing, or analysis, and can be operated remotely. To get remote access first e-mail Kari, then instructions for remote workstation access can be found here.