function [stack, img_read] = tiffread(filename, img_first, img_last) % tiffread, version 2.21b % % [stack, nbImages] = tiffread; % [stack, nbImages] = tiffread(filename); % [stack, nbImages] = tiffread(filename, imageIndex); % [stack, nbImages] = tiffread(filename, firstImageIndex, lastImageIndex); % % Reads 8,16,32 bits uncompressed grayscale and (some) color tiff files, % as well as stacks or multiple tiff images, for example those produced % by metamorph or NIH-image. However, the entire TIFF standard is not % supported (but you may extend it). % % The function can be called with a file name in the current directory, % or without argument, in which case it pop up a file openning dialog % to allow manual selection of the file. % If the stacks contains multiples images, loading can be restricted by % specifying the first and last images to read, or just one image to read. % % at return, nbimages contains the number of images read, and S is a vector % containing the different images with some additional informations. The % image pixels values are stored in the field .data, for gray level images, % or in the fields .red, .green and .blue % the pixels values are in the native (integer) format, % and must be converted to be used in most matlab functions. % % Example: % im = tiffread('spindle.stk'); % imshow( double(im(5).data), [] ); % % Francois Nedelec, EMBL, Copyright 1999-2006. % rewriten July 7th, 2004 at Woods Hole during the physiology course. % last modified Jan. 4th 2006. % Contributions: % Kendra Burbank suggested the waitbar % Hidenao Iwai for the code to read floating point images, % Stephen Lang made tiffread more compliant with PlanarConfiguration % Elias Beauchamp to read Metamorph camera info % % Please, help us improve this software: send us feedback/bugs/suggestions % This software is provided at no cost by a public research institution. % % Francois Nedelec % nedelec (at) embl.de % Cell Biology and Biophysics, EMBL; Meyerhofstrasse 1; 69117 Heidelberg; Germany % http://www.embl.org % http://www.cytosim.org % % Modified slightly by DK on Feb 12/07 to recognize MetaMorph 'Offset' % and 'TriggerOn' parameters, and extract pixelsize and timestep info from % MM STK files %Optimization: join adjacent TIF strips: this results in faster reads consolidateStrips = 1; %if there is no argument, we ask the user to choose a file: if (nargin == 0) [filename, pathname] = uigetfile('*.tif;*.stk;*.lsm', 'select image file'); filename = [ pathname, filename ]; end if (nargin<=1) img_first = 1; img_last = 10000; end if (nargin==2) img_last = img_first; end % not all valid tiff tags have been included, as they are really a lot... % if needed, tags can easily be added to this code % See the official list of tags: % http://partners.adobe.com/asn/developer/pdfs/tn/TIFF6.pdf % % the structure IMG is returned to the user, while TIF is not. % so tags usefull to the user should be stored as fields in IMG, while % those used only internally can be stored in TIF. global TIF; TIF = []; %counters for the number of images read and skipped img_skip = 0; img_read = 0; % set defaults values : TIF.SampleFormat = 1; TIF.SamplesPerPixel = 1; TIF.BOS = 'l'; %byte order string if isempty(findstr(filename,'.')) filename = [filename,'.tif']; end [TIF.file, message] = fopen(filename,'r','l'); if TIF.file == -1 filename = strrep(filename, '.tif', '.stk'); [TIF.file, message] = fopen(filename,'r','l'); if TIF.file == -1 error(['file <',filename,'> not found.']); end end % read header % read byte order: II = little endian, MM = big endian byte_order = setstr(fread(TIF.file, 2, 'uchar')); if ( strcmp(byte_order', 'II') ) TIF.BOS = 'l'; %normal PC format elseif ( strcmp(byte_order','MM') ) TIF.BOS = 'b'; else error('This is not a TIFF file (no MM or II).'); end %----- read in a number which identifies file as TIFF format tiff_id = fread(TIF.file,1,'uint16', TIF.BOS); if (tiff_id ~= 42) error('This is not a TIFF file (missing 42).'); end %----- read the byte offset for the first image file directory (IFD) ifd_pos = fread(TIF.file,1,'uint32', TIF.BOS); while (ifd_pos ~= 0) clear IMG; IMG.filename = fullfile( pwd, filename ); % move in the file to the first IFD fseek(TIF.file, ifd_pos, -1); %disp(strcat('reading img at pos :',num2str(ifd_pos))); %read in the number of IFD entries num_entries = fread(TIF.file,1,'uint16', TIF.BOS); %disp(strcat('num_entries =', num2str(num_entries))); %read and process each IFD entry for i = 1:num_entries % save the current position in the file file_pos = ftell(TIF.file); % read entry tag TIF.entry_tag = fread(TIF.file, 1, 'uint16', TIF.BOS); entry = readIFDentry; %disp(strcat('reading entry <',num2str(TIF.entry_tag),'>')); switch TIF.entry_tag case 254 TIF.NewSubfiletype = entry.val; case 256 % image width - number of column IMG.width = entry.val; case 257 % image height - number of row IMG.height = entry.val; TIF.ImageLength = entry.val; case 258 % BitsPerSample per sample TIF.BitsPerSample = entry.val; TIF.BytesPerSample = TIF.BitsPerSample / 8; IMG.bits = TIF.BitsPerSample(1); %fprintf(1,'BitsPerSample %i %i %i\n', entry.val); case 259 % compression if (entry.val ~= 1) error('Compression format not supported.'); end case 262 % photometric interpretation TIF.PhotometricInterpretation = entry.val; if ( TIF.PhotometricInterpretation == 3 ) fprintf(1, 'warning: ignoring the look-up table defined in the TIFF file'); end case 269 IMG.document_name = entry.val; case 270 % comment: try TIF.info = parseMM_info(entry.val); catch warning('Unable to parse MM info.') end case 271 IMG.make = entry.val; case 273 % strip offset TIF.StripOffsets = entry.val; TIF.StripNumber = entry.cnt; %fprintf(1,'StripNumber = %i, size(StripOffsets) = %i %i\n', TIF.StripNumber, size(TIF.StripOffsets)); case 277 % sample_per pixel TIF.SamplesPerPixel = entry.val; %fprintf(1,'Color image: sample_per_pixel=%i\n', TIF.SamplesPerPixel); case 278 % rows per strip TIF.RowsPerStrip = entry.val; case 279 % strip byte counts - number of bytes in each strip after any compressio TIF.StripByteCounts= entry.val; case 282 % X resolution IMG.x_resolution = entry.val; case 283 % Y resolution IMG.y_resolution = entry.val; case 284 %planar configuration describe the order of RGB TIF.PlanarConfiguration = entry.val; case 296 % resolution unit IMG.resolution_unit= entry.val; case 305 % software IMG.software = entry.val; case 306 % datetime IMG.datetime = entry.val; case 315 IMG.artist = entry.val; case 317 %predictor for compression if (entry.val ~= 1) error('unsuported predictor value'); end case 320 % color map IMG.cmap = entry.val; IMG.colors = entry.cnt/3; case 339 TIF.SampleFormat = entry.val; if (( TIF.SampleFormat ~= 1) & (TIF.SampleFormat ~= 3)) error(sprintf('unsuported sample format = %i', TIF.SampleFormat)); end case 33628 %metamorph specific data IMG.MM_private1 = entry.val; case 33629 %this tag identify the image as a Metamorph stack! TIF.MM_stack = entry.val; TIF.MM_stackCnt = entry.cnt; if ( img_last > img_first ) waitbar_handle = waitbar(0,'Please wait...','Name',['Reading ' filename]); end case 33630 %metamorph stack data: wavelength TIF.MM_wavelength = entry.val; case 33631 %metamorph stack data: gain/background? TIF.MM_private2 = entry.val; case 34412 % Zeiss LSM data (I have no idea what that represents...) IMG.LSM = entry.val; otherwise fprintf(1,'ignored TIFF entry with tag %i (cnt %i)\n', TIF.entry_tag, entry.cnt); end % move to next IFD entry in the file fseek(TIF.file, file_pos+12,-1); end %Planar configuration is not fully supported %Per tiff spec 6.0 PlanarConfiguration irrelevent if SamplesPerPixel==1 %Contributed by Stephen Lang if ((TIF.SamplesPerPixel ~= 1) & (TIF.PlanarConfiguration == 1)) error(sprintf('PlanarConfiguration = %i not supported', TIF.PlanarConfiguration)); end %total number of bytes per image: PlaneBytesCnt = IMG.width * IMG.height * TIF.BytesPerSample; if consolidateStrips %Try to consolidate the strips into a single one to speed-up reading: BytesCnt = TIF.StripByteCounts(1); if BytesCnt < PlaneBytesCnt ConsolidateCnt = 1; %Count how many Strip are needed to produce a plane while TIF.StripOffsets(1) + BytesCnt == TIF.StripOffsets(ConsolidateCnt+1) ConsolidateCnt = ConsolidateCnt + 1; BytesCnt = BytesCnt + TIF.StripByteCounts(ConsolidateCnt); if ( BytesCnt >= PlaneBytesCnt ) break; end end %Consolidate the Strips if ( BytesCnt <= PlaneBytesCnt(1) ) & ( ConsolidateCnt > 1 ) %fprintf(1,'Consolidating %i stripes out of %i', ConsolidateCnt, TIF.StripNumber); TIF.StripByteCounts = [BytesCnt; TIF.StripByteCounts(ConsolidateCnt+1:TIF.StripNumber ) ]; TIF.StripOffsets = TIF.StripOffsets( [1 , ConsolidateCnt+1:TIF.StripNumber] ); TIF.StripNumber = 1 + TIF.StripNumber - ConsolidateCnt; end end end %read the next IFD address: ifd_pos = fread(TIF.file, 1, 'uint32', TIF.BOS); if (ifd_pos) disp(['next ifd at', num2str(ifd_pos)]); end if isfield( TIF, 'MM_stack' ) if ( img_last > TIF.MM_stackCnt ) img_last = TIF.MM_stackCnt; end %this loop is to read metamorph stacks: for ii = img_first:img_last TIF.StripCnt = 1; %read the image fileOffset = PlaneBytesCnt * ( ii - 1 ); %fileOffset = 0; %fileOffset = ftell(TIF.file) - TIF.StripOffsets(1); if ( TIF.SamplesPerPixel == 1 ) IMG.data = read_plane(fileOffset, IMG.width, IMG.height, 1); else IMG.red = read_plane(fileOffset, IMG.width, IMG.height, 1); IMG.green = read_plane(fileOffset, IMG.width, IMG.height, 2); IMG.blue = read_plane(fileOffset, IMG.width, IMG.height, 3); end % print a text timer on the main window, or update the waitbar % fprintf(1,'img_read %i img_skip %i\n', img_read, img_skip); try waitbar( img_read/TIF.MM_stackCnt, waitbar_handle); catch stack = 0; return end [ IMG.info, IMG.MM_stack, IMG.MM_wavelength, IMG.MM_private2 ] = extractMetamorphData(ii); img_read = img_read + 1; stack( img_read ) = IMG; end break; else %this part to read a normal TIFF stack: if ( img_skip + 1 >= img_first ) TIF.StripCnt = 1; %read the image if ( TIF.SamplesPerPixel == 1 ) IMG.data = read_plane(0, IMG.width, IMG.height, 1); else IMG.red = read_plane(0, IMG.width, IMG.height, 1); IMG.green = read_plane(0, IMG.width, IMG.height, 2); IMG.blue = read_plane(0, IMG.width, IMG.height, 3); end img_read = img_read + 1; try stack( img_read ) = IMG; catch %stack %IMG error('The file contains dissimilar images: you can only read them one by one'); end else img_skip = img_skip + 1; end if ( img_skip + img_read >= img_last ) break; end end end % Extracts the x/y pixel size using MetaMorph data fseek(TIF.file,stack(1).MM_private1(10),'bof'); pixelSizeNumAndDenom=fread(TIF.file,2,'uint32',TIF.BOS); [stack.pixelSize] = deal(pixelSizeNumAndDenom(1)/pixelSizeNumAndDenom(2)); % Extracts the time step using MetaMorph data temp = [stack.MM_stack]; time = temp(4,:); % convert ms to seconds and averages over all frames [stack.timeStep] = deal(mean(diff(time))/1000); %clean-up fclose(TIF.file); if exist('waitbar_handle', 'var') delete( waitbar_handle ); clear waitbar_handle; end drawnow; %return empty array if nothing was read if ~ exist( 'stack', 'var') stack = []; end return; %============================================================================ function plane = read_plane(offset, width, height, planeCnt ) global TIF; %return an empty array if the sample format has zero bits if ( TIF.BitsPerSample(planeCnt) == 0 ) plane=[]; return; end %fprintf(1,'reading plane %i size %i %i\n', planeCnt, width, height); % Preallocate the matrix to hold the data: %string description of the type of integer needed: int8 or int16... typecode = sprintf('int%i', TIF.BitsPerSample(planeCnt) ); %unsigned int if SampleFormat == 1 if ( TIF.SampleFormat == 1 ) typecode = [ 'u', typecode ]; % Preallocate a matrix to hold the sample data: plane = eval( [ typecode, '(zeros(width, height))'] ); elseif ( TIF.SampleFormat == 3 ) typecode = 'float'; plane = zeros(width, height); end line = 1; while ( TIF.StripCnt <= TIF.StripNumber ) strip = read_strip(offset, width, planeCnt, TIF.StripCnt, typecode ); TIF.StripCnt = TIF.StripCnt + 1; % copy the strip onto the data plane(:, line:(line+size(strip,2)-1)) = strip; line = line + size(strip,2); if ( line > height ) break; end end % Extract valid part of data if needed if ~all(size(plane) == [width height]), plane = plane(1:width, 1:height); error('Cropping data: more bytes read than needed...'); end % transpose the image plane = plane'; return; %=================== sub-functions to read a strip =================== function strip = read_strip(offset, width, planeCnt, stripCnt, typecode) global TIF; %fprintf(1,'reading strip at position %i\n',TIF.StripOffsets(stripCnt) + offset); StripLength = TIF.StripByteCounts(stripCnt) ./ TIF.BytesPerSample(planeCnt); %fprintf(1, 'reading strip %i\n', stripCnt); fseek(TIF.file, TIF.StripOffsets(stripCnt) + offset, 'bof'); bytes = fread( TIF.file, StripLength, typecode, TIF.BOS ); if ( length(bytes) ~= StripLength ) error('End of file reached unexpectedly.'); end strip = reshape(bytes, width, StripLength / width); return; %===================sub-functions that reads an IFD entry:=================== function [nbbytes, typechar] = matlab_type(tiff_typecode) switch (tiff_typecode) case 1 nbbytes=1; typechar='uint8'; case 2 nbbytes=1; typechar='uchar'; case 3 nbbytes=2; typechar='uint16'; case 4 nbbytes=4; typechar='uint32'; case 5 nbbytes=8; typechar='uint32'; otherwise error('tiff type not supported') end return; %===================sub-functions that reads an IFD entry:=================== function entry = readIFDentry() global TIF; entry.typecode = fread(TIF.file, 1, 'uint16', TIF.BOS); entry.cnt = fread(TIF.file, 1, 'uint32', TIF.BOS); %disp(['typecode =', num2str(entry.typecode),', cnt = ',num2str(entry.cnt)]); [ entry.nbbytes, entry.typechar ] = matlab_type(entry.typecode); if entry.nbbytes * entry.cnt > 4 %next field contains an offset: offset = fread(TIF.file, 1, 'uint32', TIF.BOS); %disp(strcat('offset = ', num2str(offset))); fseek(TIF.file, offset, -1); end if TIF.entry_tag == 33629 %special metamorph 'rationals' entry.val = fread(TIF.file, 6*entry.cnt, entry.typechar, TIF.BOS); else if entry.typecode == 5 entry.val = fread(TIF.file, 2*entry.cnt, entry.typechar, TIF.BOS); else entry.val = fread(TIF.file, entry.cnt, entry.typechar, TIF.BOS); end end if ( entry.typecode == 2 ) entry.val = char(entry.val'); end return; %==============distribute the metamorph infos to each frame: function [info, stack, wavelength, private2 ] = extractMetamorphData(imgCnt) global TIF; info = []; stack = []; wavelength = []; private2 = []; if TIF.MM_stackCnt == 1 return; end left = imgCnt - 1; if isfield( TIF, 'info' ) % S = length(TIF.info) / TIF.MM_stackCnt; info = TIF.info(imgCnt); end if isfield( TIF, 'MM_stack' ) S = length(TIF.MM_stack) / TIF.MM_stackCnt; stack = TIF.MM_stack( [S*left+1:S*left+S] ); end if isfield( TIF, 'MM_wavelength' ) S = length(TIF.MM_wavelength) / TIF.MM_stackCnt; wavelength = TIF.MM_wavelength( [S*left+1:S*left+S] ); end if isfield( TIF, 'MM_private2' ) S = length(TIF.MM_private2) / TIF.MM_stackCnt; private2 = TIF.MM_private2( [S*left+1:S*left+S] ); end return; %%%% Parse the Metamorph camera info tag into respective fields % EVBR 2/7/2005 function mm_infor = parseMM_info(info) i=0; kk=info; kk(find(double(kk)==0)) = ''; while(length(kk)) i=i+1; [tmp, kk] = strtok(kk, 13); % [tmp,b] = strtok(tmp, 0); token{i} = sscanf(tmp, '%s'); end tok = {token{1:end-1}}; len = length(tok)/11; k=0; for i=1:length(tok) if ~isempty(token{i}) [tkk, remk] = strtok(token{i}, 58); if strcmp(tkk, 'Exposure') k=k+1; end switch tkk case 'Exposure' % Exposure: 200 ms -> 200 mm_infor(k).Exposure = str2double(remk(2:strfind(remk, 'ms')-1)); case 'Binning' % Binning: 1 x 1 -> [1 1] tmp = sscanf(remk, '%c %d %c %d')'; mm_infor(k).Binning = tmp([2,4]); case 'Region' % Region: 1392 x 1040, offset at (0, 0) -> [1392 1040], [0 0] [a,b] = strtok(remk, ','); tmp = sscanf(a, '%c %d %c %d')'; mm_infor(k).Region.Size = tmp([2,4]); [a,b] = strtok(b, ','); mm_infor(k).Region.Offset = [str2double(a(end)), str2double(b(2))]; case 'Subtract' % Subtract: Off -> 0 mm_infor(k).Subtract = ~strcmp(remk(2:end),'Off'); case 'Shading' % Shading: Off -> 0 mm_infor(k).Shading = ~strcmp(remk(2:end),'Off'); case 'Digitizer' % Digitizer: 5MHz -> 5 mm_infor(k).Digitizer = sscanf(remk(2:end), '%d %*s')'; case 'Gain' % Gain: Gain 1 (1x) -> 1 mm_infor(k).Gain = sscanf(remk(strfind(remk,'(')+1:end),... '%d %*s %d')'; case 'CameraShutter' % Camera Shutter: Always Open -> 'AlwaysOpen' mm_infor(k).CameraShutter = remk(2:end); case 'ClearCount' % Clear Count: 2 -> 2 tmp = sscanf(remk, '%c %d')'; mm_infor(k).ClearCount = tmp(2); case 'TriggerMode' % Trigger Mode: Normal -> 'Normal' mm_infor(k).TriggerMode = remk(2:end); case 'Temperature' % Temperature: 20 -> 20 tmp = sscanf(remk, '%c %d')'; mm_infor(k).Temperature = tmp(2); case 'Offset' tmp = sscanf(remk, '%c %d')'; mm_infor(k).Offset = tmp(2); case 'TriggerOn' tmp = sscanf(remk, '%c %d')'; mm_infor(k).TriggerOn = tmp(2); case 'Illumination' tmp = strtok(remk,':'); mm_infor(k).Illumination = tmp; case 'MultiDimensionsExperiment' tmp = strtok(remk,':'); mm_infor(k).MultiDimensionsExperiment = tmp; otherwise warning('Uknown MM_Tag') end end end