Method for forming an image reference device
A method is provided of forming an imaging reference device for a biological material imaging system. A first region is provided in a substrate formed from an optically transmissive material which is an analogue of biological tissue. This first region is stained using a first biological tissue stain. A second region is provided in second substrate which is optically transmissive and an analogue of biological tissue. The second region is stained with a second biological tissue stain. The first and second regions are overlapped as a pair, such that light incident upon the pair is modulated by the respective first and second biological tissue stains of the respective regions. Multiple regions of the first and second type may be provided, stained to different degrees.
1. A method of forming an imaging reference device for a biological material imaging system, the method comprising:
providing at least one first region of a first substrate, the first substrate being formed from an optically transmissive material which is an analogue of biological tissue, wherein the at least one first region is stained using a first biological tissue stain;
providing at least one second region of a second substrate, the second substrate being formed from an optically transmissive material which is an analogue of biological tissue, wherein the at least one second region is stained using a second biological tissue stain; and,
overlapping at least one pair of the regions, each pair being formed from the overlap of a first region with a second region, such that light incident upon a pair is modulated by the respective first and second biological tissue stains of the respective regions.
2. A method according to claim 1 , wherein a plurality of first regions is provided as a first series of first regions and wherein a plurality of second regions is provided as a second series of second regions.
3. A method according to claim 2 , wherein in one or each of the first and second series, the respective spectral effects of the stains applied to the regions within a series are modulated.
4. A method according to claim 3 , wherein the modulated spectral effects are effected by using different concentrations of the respective biological tissue stains.
5. A method according to claim 3 , wherein the modulated spectral effects are effected by using different time durations of application of the respective biological tissue stains.
6. A method according to claim 3 , wherein the modulated spectral effects are caused by using regions of the respective first and/or second substrate having different thicknesses.
7. A method according to claim 3 , wherein the modulated spectral effects are caused by using regions of the respective first and/or second substrate having different stain absorption characteristics.
8. A method according to claim 2 , wherein each region in one or each of the first or second series is elongate and wherein the directions of elongation of the regions are parallel.
9. A method according to claim 8 , wherein the regions of the first and the second series are elongate, wherein the respective directions of elongation of the first and second series are arranged at an angle such that the pairs of overlapping regions form a two dimensional array.
10. A method according to claim 1 , wherein one or each of the first substrate or second substrate is provided as a film.
11. A method according to claim 1 wherein the physical arrangement of the regions within the first or second series corresponds to a monotonical modulation in the effect of the respective staining on incident light.
12. A method according to claim 1 , wherein the stained regions within a series are formed by the overlap of corresponding numbers of layers of the substrate, each of similar thickness and staining.
13. A method according to claim 1 the first biological tissue stain and the second biological tissue stain comprise one or more different biological tissue stains.
14. A method according to claim 1 wherein the first and second stains are the stains used for one of the following staining protocols:
Haematoxylin and Eosin (H&E), Diaminobenzidine (DAB) with Haematoxylin counter stain, Papanicolaou (PAP), Perls' Prussian blue, Periodic acid-Schiff (PAS), Reticulin, Millers elastic Van Gieson, Shikata, Giemsa stain, Ziehl Neelsen technique, Grocott, Alcian blue PAS, Jones methenamine silver, Gram, Congo red stain for amyloid and Masson trichrome.
15. A method according to claim 1 , wherein the one or each of the first and second substrates is formed from a material selected from the group comprising: cellophane, cellulose, agar, agarose and gelatine.
16. A method according to claim 1 , further comprising attaching the device to a microscope slide.
17. A method according to claim 1 , wherein the staining is provided by immersing the respective first or second substrate in a bath of stain material having a predetermined composition and for a predetermined period.
18. An imaging reference device for a biological material imaging system, the device comprising:
a first substrate formed from an optically transmissive material which is an analogue of biological tissue, the first substrate having at least one first region stained using a first biological tissue stain;
a second substrate formed from an optically transmissive material which is an analogue of biological tissue, the second substrate having at least one second region stained using a first biological tissue stain;
wherein at least one of the first and second regions is overlapped so as to form a pair of regions, such that light incident upon a pair is modulated by the respective first and second biological tissue stains of the respective regions.
19. An imaging reference device according to claim 18 , wherein the first and second substrates are connected to each other.
20. An imaging reference device according to claim 18 , wherein the device is a calibration device for a biological imaging system.