This document contains a portfolio of images taken by Fred Martin for various biology projects. It includes over 50 microscope images of specimens such as C. elegans, plant tissues, cells and buccal cells. The images were taken using different microscopes, filters, objectives and techniques such as brightfield, fluorescence, confocal, DIC and stitching. Accompanying each image is metadata on the microscope, specimen, technique and settings used.

1. PROJECT PORTFOLIO FRED MARTIN 5/08/2010 Dr. Giorgi BIOSC 3

2. C. elegans on Nikon 80i, DIC 20x DIC, Bin 1x1, 8 bit, Exp = 10.0ms, Black 495 White 4095 gamma 1.0

3. e BF Nikon 80i Yucca Stem 20 f BF Nikon 80i Cerebellum, human 19 h Phase Olympus Drospophilia Wing 18 g,f BF Olympus Corn Root Tip 17 a,d DIC Zeiss FluoCells prepared slide #4: Intestine 16 g BF, DAPI LP filter Leica BF Pseudo Stratified Ciliated Columnar Epithelial 15 g BF Leica BF Pseudo Stratified Ciliated Columnar Epithelial 14 g BF Nikon 80i Brine Shrimp 13 g BF Nikon 80i Lycodopia pollen 12 e BF, RL Olympus Abutilon x hybridium 11 a RL Nikon 80i Lypha Root Tip 10 a,d RL + DIC Nikon 80i Lily Anther 9 a RL Nikon 80i Lily Anther 8 a,b RL Zeiss FluoCells® prepared slide #1: BPAEC 7 a,b RL Zeiss FluoCells® prepared slide #1: BPAEC 6 a,b RL Zeiss FluoCells prepared slide #2: BPAE 5 a,b RL Nikon 80i FluoCells prepared slide #4: Intestine 4 a,b RL Nikon 80i FluoCells prepared slide #4: Intestine 3 a,d RL + DIC Nikon 80i C. elegans 2 d DIC Nikon 80i C. elegans 1 Requirement Technique Scope Specimen Slide # Project Image Index

4. Image Index Project Slide # Specimen Scope Technique Requmt b,f RL Nikon 80i FluoCells prepared slide #2: BPAE 38 i Confocal SP5 FluoCells® prepared slide #6: Muntjac Fibroblast 37 c Confocal SP5 FluoCells® prepared slide #6: Muntjac Fibroblast 36 c Confocal SP5 FluoCells® prepared slide #6: Muntjac Fibroblast 35 c,j Confocal SD Lily Anther pollen 34 c Confocal SD Lily Anther pollen 33 c Confocal SD Lily Anther pollen 32 c Confocal SP5 C. elegans 31 c Confocal Zeiss710 C. elegans 30 c Confocal Zeiss710 C. elegans 29 c Confocal Zeiss710 C. elegans 28 l RL Nikon 80i C. elegans 27 l RL Nikon 80i C. elegans 26 j RL Nikon 80i C. elegans 25 c,h Confocal SP5 Buccal cells 24 c,h Confocal SP5 Buccal cells 23 c,h Confocal SP5 Buccal cells 22 d DIC Nikon 80i C. elegans 21

5. Project SS: C. elegans on Nikon 80i, 3 channel RL + DIC 20x DIC, Bin 1x1, 8 bit, Exp = 10.0ms, Black 495 White 4095 gamma 1.0 DAPI 2.59s, FITC 9.76s, TRITC 4.9s

6. Invt4_10x_CompositeRGB_002.jpg 20091207 /Nikon2 /10x ; DIC: 73ms/ 1,503/ 0.95/ 4,095 R: 97ms/ 79/ 0.63/ 4,095 G: 200ms/ 239/ 0.95/4,095 B:500ms/223/0.77/4,095 SPECIMEN (A1) NIKON, FLUORESENCE, MOLECULAR PROBES SLIDE: TISSUE

7. invt4_a1_Composite_RG_DapiLUT_75.jpg 20091123 /Nikon2 /20x ; DIC: 20ms/ 79/ 0.94/ 3,310 R: 47ms/ 79/ 0.94/ 3,310 G: 41ms/ 95/ 0.67/3,054 B:392ms/159/0.51/2,830 SPECIMEN (A2) NIKON, FLUORESENCE, MOLECULAR PROBES SLIDE: TISSUE

8. FluoCells prepared slide #2: BPAE bovine pulmonary artery endothelial cells Image A - 20091105: Zeiss #1 20x ZEISS, FLUORESENCE, MOLECULAR PROBES SLIDE: CELLS

9. FluoCells ® prepared slide #1: BPAEC, Zeiss 63x 20091016_INVG1_AREA3_CompositeAdj_jpg.jpg ZEISS, FLUORESENCE, MOLECULAR PROBES SLIDE: CELLS

10. NIKON, FLUORESENCE, MOLECULAR PROBES SLIDE: CELLS FM_INVT1_A1_CompositeAdjRGB_xDIC BPAE CELLS INVITROGEN #1

11. 20x_LilyAnther_Fluro_RGB.jpg 20091123 /Nikon2 /20x ; R: 47ms/ 79/ 0.94/ 3,310 G: 41ms/ 95/ 0.67/3,054 B:392ms/159/0.51/2,830 SPECIMEN (B) NIKON, FLUORESENCE

12. FM_lycopodiumCaptured_FluoroRGBnDIC_20x_2.jpg 20091123 /Nikon2 /20x ; DIC: 20ms/ 79/ 0.94/ 3,310 R: 47ms/ 79/ 0.94/ 3,310 G: 41ms/ 95/ 0.67/3,054 B:392ms/159/0.51/2,830 SPECIMEN (C) NIKON, FLUORESENCE + DIC

13. SPECIMEN (F) Typha_10x_Composite_001_RGB_Adj 20091207 /Nikon2 /10x ; DIC: 73ms/ 1,503/ 0.95/ 4,095 R: 97ms/ 79/ 0.63/ 4,095 G: 200ms/ 239/ 0.95/4,095 B:500ms/223/0.77/4,095 COMPOSITE CHANNEL: RED CHANNEL: GREEN CHANNEL: BLUE NIKON, FLUORESCENSE (3 CHANNELS + OVERLAY)

14. Pollen Grain: Abutilon x hybridium @ 20x EM Channel: Green Exp: 96.7ms Gain:7.1 Offset: -876 EM Channel: Red Exp: 96.7ms Gain:7.1 Offset: -876 EM Channel: (BF) Exp: 16.7ms Gain: 4.5 Offset: -161 Composite OLYMPUS, OWN SPECIMEN, FLUORO OVERLAY + 3 CHANNELS

15. fm__pseudocolor_lycopod_20x_CompositeRGBnDIC.jpg SPECIMEN (D) 20091207 /Nikon2 /20x ; “R”: 13ms/ 159/ 0.51/ 2,830 “G”: 6.6ms/ 159/ 0.51/ 2,830 “B”: 12.0ms/ 159/ 0.51/ 2,830 NIKON, BRIGHTFIELD (3 CHANNEL PSEUDO-COLORIZATION)

16. Multichannel _Shrimp_Pseudocolor2_RGB.jpg SPECIMEN (E) 20091118 /Nikon4 /20x ; R: --ms/ --/ --/ -- G: --ms/ --/ --/ -- --B: ms/ --/ --/ -- Note: metadata not logged NIKON, BRIGHTFIELD (3 CHANNEL PSEUDO-COLORIZATION)

17. Cilia PSCCE Cells Basal Lamina Pseudo Stratified Ciliated Columnar Epithelial Cross section: area 3 image Cilia Basal Lamina Nucleus Cilia (Apparent Stratification) Exp. 500.7ms Hist: 79/252 Gamma: 0.99 Auto_exp: OFF Gain: 3.1x 2009.10.01 Leica #2 60x (dry) Nucleolus { } LEICA, BRIGHTFIELD

18. PSCCE Cells Cell Membrane Cilia (Apparent Stratification) Nuclei Basal Lamina LEICA, BRIGHTFIELD 48 BitDepth: 20 / 255 Histogram: On Auto_Exp? 0.97 Gamma: 2009-10-07 DateImage: 53 % Brightness: 2.2 x Gain: 1.0 s Exposure:

19. ZEISS, DIC FM_INVT4_A1_DIC-0005.JPG 20091209 /ZEISS 1 /20x ; DIC : EXP 69ms/ CT -0.50 / GAMMA 1.0 / BRT 1.0

20. Interphase Metaphase Anaphase Telophase Prophase RESULTS: DIGITAL COUNT DETAIL – FIELD OF VIEW A: Onion Root Tip Meta Data Scope: Olympus 3 (A) 10x, rheo: full Iris ~ 30% 1.1 ms 2.8 gain -1,860 offset Note: Manual count done at 20x on Olympus3 OLYMPUS, MORPHOMETRY

21. OLYMPUS, OWN SPECIMEN, Phase Contrast

22. 4x 10x 20x Area Hunt B : Same Field of View at different magnifications: comparison of gross tissue structure/features and unique “target” reference point (marked with * ) * * * * These different Field of View images are representative of the reference sketches I made…

23. Project Marvelous Motorization: Montage (Yucca Stem c.s.) denotes stitch line Montage is 6 x 4 frames Red Nikon, 4x, exp. = 750 ų s, B/W level: 35/3900

24. Project Marvelous Motorization: Montage (C. elegans) denotes stitch line Montage is 2 x 1 frames Red Nikon, 20x, DIC w/ ¼ wave plate in, exp. = 10ms, B/W level: 495/4095

25. Own Buccal Cells cells stained with ER Tracker, : max projection of 66 slice Z-stack Leica SP5 Confocal: Lily Anther Pollen, FITC channel

26. Buccal Cells Series013 – ER Tracker, Max Projection 54 slice Z-stck, 3.7 zoom factor Note: peri-nuclear staining differences Peri-nuclear membrane staining with apparent channels / invaginations showing

27. Buccal Cells – ER Tracker, Max Projection 33 slice Z-stck Note: peri-nuclear staining differences Dead cells? RER? SER?

28. Z stack: slice series (C. elegans) Nikon, 63x oil, Z-series: 21steps @ 0.6ų/step DAPI channel exp. = 1s, B/W level: 123/3013

29. Z stack: Max. Projection (C. elegans) Nikon , 63x oil, Projection of Z-series: 21steps @ 0.6ų/step DAPI channel exp. = 1s, B/W level: 123/3013

30. Z stack: 3D surface representation (C. elegans) Nikon , 63x oil, 3d rep. of Z-series: 21steps @ 0.6ų/step DAPI channel exp. = 1s, B/W level: 123/3013

31. C. elegans on Zeiss 710 (63x oil, Max projection – 32 slice Area showing meitotic divisions in sperm formation: capsules around sperm were the structure of interest in the study)

32. Project SS: C. elegans on Zeiss 710 (63x oil, max projection 17 slice) Area near tail of male c. elegans showing spicuola

33. CONFOCAL: C. elegans on Zeiss 710 (63x oil)

34. C. elegans on Leica SP5 Slice 37 of 87 from z stack (MetaData at end of presentation)

35. Leica SD Confocal: Lily Anther Pollen, Max projection overlay (eGFP,TxRed)

36. Leica SD Confocal: Lily Anther Pollen, eGFP channel: max projection

37. Project SD: Lily Anther Pollen, view of all Z slices: screen shot

38. Frame 1 @ 0.33sec Frame 2 @ 0.67sec Frame 3 @ 1.00sec Frame 4 @ 1.33sec Frame 5 @ 1.67sec Frame 6 @ 2.00sec Frame 7 @ 2.33sec Frame 8 @ 2.67sec Frame 9 @ 3.00sec Frame 10 @ 3.33sec Frame | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | Lambda Scan from 550-610nm: Image Series above correlate to each wavelength/intensity reading in chart/graph to right Structure is fibroblast actin labeled with Alexa Fluor 488: weaker, post-peak EM signal

39. Original Separation 1 Separation 2 Dye Separation Series: 3 channels, Auto- separate, weak strength Original

40. SP5 Confocal: Mark/Find Function: 3 random x,y,z coordinates identified & recaptured later Top row series: manual images before defining xyz positions with “Mark” function Bottom row series: Auto images captured with “Find” function by recalled xyz positions

41. Morphometry Project: Comparison of Manual counts in NIS Elements vs Image Pro DAPI Channel Manual Counts Vs Image Pro Variance Comparison FOV 1 FOV 2 FOV 3 FOV 4 FOV 1 count = 27 FOV 2 count = 18 FOV 3 count = 15 (net13) FOV 4 count = 14 Counted as 3 vs 1 (+2) Counted as 1 vs 0 (+1) Counted as 1 vs 2 <same> Not Counted vs 2 (+2)