CryoSPARC Live v5.0.4 shows "No Exposures" and cannot import TIFF movie files

CryoSPARC Live
1

Hello CryoSPARC team,

I am trying to process a Cryo-EM dataset collected at PNCC on a Titan Krios 300 kV microscope. The raw data are stored as .tiff movie files.

System Information

  • CryoSPARC version: v5.0.4

  • Microscope: Titan Krios 300 kV

  • Raw pixel size: 0.4125 Å

  • Cs: 2.7 mm

  • Total dose: 41 e⁻/Ų

Data Files

The movie files are located in:

/path/to/movies/

Example filenames:

FoilHole_xxxxx_Data_xxxxx_x_YYYYMMDD_HHMMSS_fractions.tiff

The directory contains only .tiff files. I do not have accompanying .xml metadata files.

The files are visible from the CryoSPARC server:

ls /path/to/movies/*.tiff | head

returns the expected movie files.

TIFF Header Information

Running IMOD header on one movie gives:

This is a TIFF file (in strips of 11520 x 682)

Number of columns, rows, sections … 11520 8184 64

Map mode … 0 (byte)

Pixel spacing (Angstroms)… 1.000 1.000 1.000

This appears to be a 64-frame movie stack rather than a single summed image.

CryoSPARC Live Issue

In CryoSPARC Live:

  • Directory to watch:

  • /path/to/movies/

  • File name wildcard filter:

  • *.tiff

  • Search recursively: enabled

The exposure group is enabled, but CryoSPARC Live reports:

Exposure Group 1 (0 found)

No Exposures

even though the directory contains many TIFF movie files.

Import Movies Issue

I also tried using a standard Import Movies job.

Initially I accidentally specified the directory itself and received:

AssertionError: Cannot import file of type /path/to/movies

After correcting the path to:

/path/to/movies/*.tiff

I still cannot successfully import the movies.

Additional Information

  • The dataset contains only .tiff movie files.

  • No .xml metadata files are available.

  • The files are accessible from the CryoSPARC server.

  • The movies appear to contain 64 frames based on the IMOD header output.

Questions

  1. Does CryoSPARC v5.0.4 support this TIFF format?

  2. Are there known TIFF formats (e.g., Thermo Fisher/Falcon TIFF movies) that require additional metadata files?

  3. Can CryoSPARC Live detect existing TIFF files in a directory, or only newly appearing files?

  4. Are the missing XML files required for movie import?

  5. Are there additional diagnostics I can run to determine whether the TIFF format itself is unsupported?

Any suggestions would be greatly appreciated.

Thank you!

2

Welcome to the forum @Jack.

Is this problem specific to CryoSPARC Live? What happens when you try to import the movies with a non-Live Import Movies job with Movies data path specified as /path/to/movies/*.tiff?