This page last changed on Mar 14, 2017 by brian.

Before you Start

  1. You'll need Java 8 (or better) installed on your computer. Most likely your computer already has this installed. But if not, got to http://java.com.
  2. Download the jar file to your computer.
  3. Double-click the jar to run the app.

You can also get the automated image analysis application here

Automated Image Analysis.

Just isn't there yet. That's why I developed the user interface below. Notes on what's been explored can be found here.

I'm working on a new method. Documentation is here

How to use it

Setup

Configure your settings. Some settings, such as the directory you want to save data into, are preserved between sessions.

Selection

Select the image you want to analyze along with the corresponding spot-definition (aka GAL) file. The application knows how to extract a single filter from a GAL that defines multiple filters. However, it assumes that all filters defined in a GAL are essentially identical.

Alignment

When you select an image and it's corresponding GAL, a new window will pop up overlaying the expected spots onto your image. Your job is to align the spots in the GAL with those on the image. Besides, rotation, scaling and translation of the spots, you can also increase the spot search area (aka diameter).

Alignment can be done directly on the image or using the sliders on the application

Aligning using Image Controls

Aligning using Slider Controls

Results

When you press the Analyze Image button, 2 data products are created:

  1. An image with the mean intensity values overlaid
  2. A tab-delimited text file with intensity statistics.

Each file is named after the image being analyzed. In addition, the filename has the UTC date that the sample was analyzed appended. For example, analyzing an image named filter_3.tif, will create files like filter_3-20090430221134.png and filter_3-20090430221134.txt.

How is the Intensity Calculated?

The analysis results are based on:

  1. the spot diameter
    • The spot diameter defines the area that will be examined for a particular spot.
  2. the block size.
    • Defines the width and height of the area to be averaged for mean intensity.
    • For example, a block size of 3 will calculate the mean for a 3 pixel by 3 pixel block.

A spot is analyzed as follows:

  1. The mean intensity for every block (defined by block size) that fits in the circle defined by spot diameter is calculated. This is done separately for each spot.
  2. The highest mean intensity value for a given spot is selected.
  3. Statistics are calculate based on the location of the selected block
  4. The location and size of the selected block is overlaid onto the results image so you can view exactly which pixels the mean intensity values are calculated from.

Image Result

Data Result

SpotNumber	SourceWell	Area	Mean	Median	Std	Min	Max
1	M-1	9	49355.0	48604	3539.1	43383	54869
2	M-1	9	45450.4	45237	3079.1	40195	49597
9	A-1	9	5662.1	5543	276.5	5327	6037
10	A-1	9	5863.2	5859	276.8	5451	6288
11	M-1	9	46826.3	47433	3527.7	40002	51593
12	M-1	9	41673.3	41311	5130.4	30775	47634
13	C-1	9	5684.9	5703	105.0	5530	5827
14	C-1	9	5766.2	5758	134.0	5548	6039
33	A-1	9	5564.0	5639	214.3	5294	5845
34	A-1	9	5945.6	5931	188.0	5671	6170
35	A-1	9	6444.4	6452	246.2	6127	6893
36	C-1	9	5225.2	5162	207.0	4906	5492
37	C-1	9	5627.9	5539	330.1	5185	6376
38	C-1	9	5669.4	5615	231.1	5375	6160
57	M-1	9	48217.7	47461	6614.6	35229	55771
58	M-1	9	49192.3	49033	6575.7	35924	57107
59	A-1	9	6161.3	6159	190.3	5933	6543
60	A-1	9	5979.8	6066	345.0	5545	6597
61	A-1	9	7102.6	6759	1233.1	5967	9732
62	C-1	9	5487.8	5491	100.7	5355	5626
63	C-1	9	5570.3	5510	176.3	5255	5807
64	C-1	9	5459.4	5419	133.5	5235	5641
89	M-1	9	46842.1	48813	5160.4	37888	52613
90	M-1	9	47047.8	47124	4265.9	38956	52583
91	A-1	9	6272.9	6315	439.5	5284	6872
92	A-1	9	6121.4	6141	103.7	5941	6317
93	C-1	9	5494.0	5515	174.1	5303	5767
94	C-1	9	4985.1	4965	124.4	4803	5149
113	M-1	9	48944.3	49991	3707.7	42914	53405
114	I-1	9	25423.6	25613	1422.0	22955	27031
115	I-1	9	25197.3	25670	988.3	23421	26450
116	I-1	9	22132.4	22443	1272.5	19932	23443
117	K-1	9	51498.3	51759	2560.4	47191	55187
118	K-1	9	51888.1	52335	1953.1	49773	55666
119	K-1	9	52646.9	52985	2061.7	49485	55796
120	M-1	9	30787.3	30075	3475.8	23921	35206
121	M-1	9	34149.1	33959	2733.5	29768	37348
149	M-1	9	43475.1	43663	3603.0	38100	48773
150	M-1	9	44489.3	44609	3332.0	39065	49303
151	I-1	9	23479.1	23175	1384.3	20487	25171
152	I-1	9	24453.8	24841	1268.0	22381	25655
153	I-1	9	23526.8	23690	1187.2	21538	24913
154	I-1	9	21597.7	21709	1017.7	19604	22678
155	K-1	9	49745.0	50732	2936.6	44501	52759
156	K-1	9	47913.0	48388	2031.7	43423	50437
157	K-1	9	51569.7	51321	2375.2	48169	55215
158	K-1	9	52897.6	52851	2552.0	47967	56047
189	M-1	9	47880.7	49051	2907.6	44013	53047
190	I-1	9	24703.0	24444	708.3	23487	25453
191	I-1	9	23219.3	23389	1014.5	21703	24619
192	I-1	9	21817.2	22028	1010.8	19986	23403
193	K-1	9	54369.1	54732	3718.3	46532	58461
194	K-1	9	50194.6	50399	2553.9	45703	53492
195	K-1	9	45502.8	44757	2679.8	41538	48237
196	M-1	9	26627.3	27296	2518.6	22277	28987
197	M-1	9	31814.9	32016	1982.2	28295	34746
225	M-1	9	42895.4	43709	4152.5	34537	47623
226	M-1	9	39661.4	39303	3612.6	33975	44311
227	G-1	9	5144.4	5127	221.5	4734	5587
228	G-1	9	5070.7	5139	298.5	4457	5575
229	E-1	9	4945.2	4971	133.0	4764	5193
230	E-1	9	5207.7	5205	87.9	5085	5343
249	M-1	9	41336.3	42004	2850.3	35625	45492
250	M-1	9	37665.2	38325	2081.2	34699	40801
251	G-1	9	5331.7	5387	138.8	5093	5550
252	G-1	9	5594.6	5527	147.4	5398	5792
253	G-1	9	5012.8	5011	125.8	4869	5259
254	E-1	9	5017.9	5034	129.5	4703	5139
255	E-1	9	5810.6	5767	466.6	5395	6926
256	E-1	9	5415.9	5379	171.3	5195	5705
281	G-1	9	4841.2	4835	145.3	4635	5095
282	G-1	9	5204.3	5143	146.4	5015	5403
283	G-1	9	5278.7	5249	271.8	4974	5822
284	E-1	9	5187.6	5143	135.8	5083	5519
285	E-1	9	5257.4	5282	66.5	5113	5323
286	E-1	9	4652.3	4648	151.5	4414	4918
305	G-1	9	4980.6	4964	168.7	4751	5307
306	G-1	9	4723.1	4763	82.8	4591	4804
307	M-1	9	27011.7	27094	1546.5	23564	28924
308	M-1	9	27658.3	28069	1809.2	24951	29929
309	E-1	9	4644.2	4662	80.3	4523	4731
310	E-1	9	4403.9	4429	134.1	4148	4559
329	M-1	9	33267.1	32311	3910.5	25015	36996
330	M-1	9	31966.2	33229	3090.1	26878	35366


Document generated by Confluence on Feb 03, 2026 14:16