REL 2023-02-03 2024-08-26 2024-08-26 0000-0001-6413-9835 Morgan Beeby Imperial College London South Kensington London Greater London United Kingdom SW7 2AZ 0000-0001-6413-9835 Morgan Beeby Imperial College London South Kensington London Greater London United Kingdom SW7 2AZ Beeby M Cohen EJ Drobnič T Nans A Rosenthal PB Medical Research Council (MRC, United Kingdom) MR/V000799/1 United Kingdom Human Frontier Science Program (HFSP) RGP0028/2021-HOCHBERG France Biotechnology and Biological Sciences Research Council (BBSRC) BB/M011178/1 United Kingdom Cancer Research UK FC001143 United Kingdom Medical Research Council (MRC, United Kingdom) FC001143 United Kingdom Wellcome Trust FC001143 United Kingdom 25.5 10.6019/EMPIAR-11580 EMDB cell EMD-16723 EMD-16724 Beeby M Drobnič T Dataset 2 (“K2”) /data/BigKahuna/Movies/2020-10-01_K2_dataset micrographs - multiframe MRC MRC 43006 32 UNSIGNED BYTE 3838 2.2 3710 2.2

Campylobacter_K2_20201001_151628_EPU 64,000 np EFTEM 2.2 A/pix 20eV slit Spot 7 1.50 um beam size 17 s exp 32 frames Dose rate: 14 e/p/s, 2.89 e/A2/s 50 e/A2 total dose 1.53 e/A2/frame -1.5 um to -3.0 um defocus Autofocus at 105,000 nP (1.05 um beam size) 0.6/1 quantifoil grids To detect holes in EPU, use 9mm defocus in GridSq mode and "allow dark holes" One shot per hole

Dataset 1 (“F3”) /data/BigKahuna/Movies/2020-06-11_Falcon3_dataset micrographs - multiframe MRC MRC 8775 10 SIGNED 16 BIT INTEGER 4096 1.748 4096 1.748

campylobacter_mini_20200611_131510_EPU 47,000 nP 1.75 A/pix 50um C2 100um obj spot 4 beam size: 1.3um defocus range: -1.5 to -3.0 1.02 s exposure 10 movie frames dose rate: 153 e/p/s total dose: 50 e/A2 dose per movie frame: 5 e/A2/frame