Command-line tool#

nima#

Analyze multichannel (default:[“G”, “R”, “C”]) tiff time-lapse stack.

TIFFSTK : Image file.

CHANNELS : Channel names.

Save: (1) representation of image channels and segmentation BN_dim.png, (2) plot of ratios and channel intensities for each label and bg vs. time BN_meas.png, (3) table of bg values */bg.csv, (4) representation of bg image and histogram at all time points for each channel BN/bg-[C1,C2,⋯]-method.pdf, and for each label: (5) table of ratios and measured properties BN/label[1,2,⋯].csv and (6) ratio images BN/label[1,2,⋯]_r[cl,pH].tif.

nima [OPTIONS] TIFFSTK [CHANNELS]...

Options

--version#

Show the version and exit.

--silent#

Do not print; verbose=0.

-o, --output <output>#

Output path [default: ./nima/].

--hotpixels#

Median filter (rad=0.5) to remove hotpixels.

-f, --flat <flat_f>#

Dark for shading correction.

-d, --dark <dark_f>#

Flat for shading correction.

--bg-method <bg_method>#

Background estimation algorithm [default:li_adaptive].

Options:

li_adaptive | entropy | arcsinh | adaptive | li_li

--bg-downscale <bg_downscale>#

Binning Y X.

--bg-radius <bg_radius>#

Radius for entropy or arcsinh methods [def:10].

--bg-adaptive-radius <bg_adaptive_radius>#

Radius for adaptive methods [def:X/2].

--bg-percentile <bg_percentile>#

Percentile for entropy or arcsinh methods [def:10].

--bg-percentile-filter <bg_percentile_filter>#

Percentile filter for arcsinh method [def:80].

--fg-method <fg_method>#

Segmentation algorithm [default:yen].

Options:

yen | li

--min-size <min_size>#

Minimum size labeled objects [def:2000].

--clear-border#

Remove labels touching image borders.

--wiener#

Wiener filter before segmentation.

--watershed#

Watershed binary mask (to label cells).

--randomwalk#

Randomwalk binary mask (to label cells).

--image-ratios, --no-image-ratios#

Compute ratio images? [default:True]

--ratio-median-radii <ratio_median_radii>#

Median filter ratio images with radii [def:(7,3)].

--channels-cl <channels_cl>#

Channels for Cl ratio [default:C/R].

--channels-ph <channels_ph>#

Channels for pH ratio [default:G/C].

Arguments

TIFFSTK#

Required argument

CHANNELS#

Optional argument(s)

bias#

Compute bias, dark and flat.

bias [OPTIONS] COMMAND [ARGS]...

Options

--version#

Show the version and exit.

-o, --output <output>#

Output path [default: *.tif, *.png].

bias#

Compute BIAS frame and estimate read noise.

FPATH: the bias stack (Light Off - 0 acquisition time).

Output:

  • .tif BIAS image = median projection

  • .png plot (histograms, median, projection, hot pixels)

  • [.csv coordinates and values of hot pixels] if detected

bias bias [OPTIONS] FPATH

Arguments

FPATH#

Required argument

dark#

Compute DARK.

FPATH: the bias stack (Light Off - Long acquisition time).

bias dark [OPTIONS] FPATH

Options

--bias <bias>#
--time <time>#

Arguments

FPATH#

Required argument

flat#

Flat from (.tf8) file.

bias flat [OPTIONS] FPATH

Options

--bias <bias>#

Arguments

FPATH#

Required argument

mflat#

Flat from a collection of (.tif) files.

bias mflat [OPTIONS] GLOBPATH

Options

--bias <bias>#

Arguments

GLOBPATH#

Required argument

plot#

Plot profiles of 2D (Bias-Flat) image.

bias plot [OPTIONS] FPATH

Arguments

FPATH#

Required argument