%PDF-1.2 % 8 0 obj << /Length 9 0 R >> stream 0.24 w 2 J BT /F1 11.52 Tf 507.732 744.257 TD (1)Tj ET BT /F2 19.2 Tf 156.844 701.296 TD 0 Tc 0 Tw (Making and Staining a Blood Smear)Tj ET BT /F1 11.52 Tf 98.762 667.455 TD (A well-made blood smear is a beauty to behold, and likely to yield interesting and)Tj ET BT 98.762 651.375 TD (significant information for a research project. A poor slide is a torment. The extra time)Tj ET BT 98.762 635.535 TD (and care taken during the field season will be rewarded later when the smears must be)Tj ET BT 98.762 619.694 TD (scanned, and parasites identified and counted. Here, the methods for making and staining)Tj ET BT 98.762 603.614 TD (smears are given, as well as a list of sources for high quality slides, stain, and chemicals.)Tj ET BT 98.762 587.773 TD (Photographs showing well-made smears are shown on the website.)Tj ET BT 98.762 555.853 TD (Dried blood samples for genetic studies should always be made at the same time as the)Tj ET BT 98.762 540.012 TD (smears. The method is very easy and modern research must combine studies of)Tj ET BT 98.762 524.172 TD (morphology under the microscope with molecular methods. The technique for making)Tj ET BT 98.762 508.332 TD (and storing dried blood samples is given in the section \322Dried Blood Samples\323.)Tj ET BT /F2 11.52 Tf 98.762 476.411 TD (Making a smear)Tj ET BT /F1 11.52 Tf 98.762 444.49 TD (1.)Tj /F3 11.52 Tf 8.64 0 TD ( )Tj /F1 11.52 Tf 8.64 0 TD (A single smear can be made per slide \(smear running the length of the slide\) or two)Tj ET BT 116.043 428.65 TD (\(or even three\) smears can share a slide, with the smears running the width of the)Tj ET BT 116.043 412.809 TD (slide. Putting two smears per slide saves on weight \(glass is heavy\) for field trips,)Tj ET BT 116.043 396.729 TD (and storage space. A picture showing both versions is included on the website.)Tj ET BT 98.762 365.048 TD (2.)Tj /F3 11.52 Tf 8.64 0 TD ( )Tj /F1 11.52 Tf 8.64 0 TD (It is easiest to use microscope slides with a frosted end, so that identifying)Tj ET BT 116.043 348.968 TD (information can be written there with pencil. Warning: Compare different pencils to)Tj ET BT 116.043 333.128 TD (find one that does not yield labels that rub off or wash off in the methanol dip.)Tj ET BT 98.762 301.207 TD (3.)Tj /F3 11.52 Tf 8.64 0 TD ( )Tj /F1 11.52 Tf 8.64 0 TD (Place a drop of blood approximately 4 mm in diameter on the slide \(near the end if)Tj ET BT 116.043 285.367 TD (one smear is to be made, or at the proper location if two smears are to share a slide\).)Tj ET BT 116.043 269.526 TD (See the drawing below.)Tj ET BT 98.762 237.605 TD (4.)Tj /F3 11.52 Tf 8.64 0 TD ( )Tj /F1 11.52 Tf 8.64 0 TD (Spread the drop by using another slide \(called here the \322spreader\323\), placing the)Tj ET BT 116.043 221.765 TD (spreader at a 45\241 angle and BACKING into the drop of blood. The spreader catches)Tj ET BT 116.043 205.685 TD (the drop and it spreads by capillary action along its edge. To make a short smear,)Tj ET BT 116.043 189.844 TD (hold the spreader at a steeper angle, and to make a longer smear, hold it closer to the)Tj ET BT 116.043 174.004 TD (drop. Now, push the spreader across the slide; this PULLS the blood across to make)Tj ET BT 116.043 157.924 TD (the smear. Do not push the blood by having it ahead of the smearing slide! It should)Tj ET BT 116.043 142.083 TD (take about one second to smear the drop. A smooth action is required, with the edge)Tj ET BT 116.043 126.243 TD (of the spreader held against the slide. This will yield a nice, even smear.)Tj ET endstream endobj 9 0 obj 3559 endobj 4 0 obj << /Type /Page /Parent 5 0 R /Resources << /Font << /F1 6 0 R /F2 7 0 R /F3 11 0 R >> /ProcSet 2 0 R >> /Contents 8 0 R >> endobj 13 0 obj << /Length 14 0 R >> stream 0.24 w BT /F1 11.52 Tf 507.732 744.257 TD (2)Tj ET 0.72 w 1 g 192.484 596.654 213.605 68.402 re f 192.124 596.294 214.325 69.122 re s 247.326 664.695 m 247.326 595.574 l S 192.484 506.652 213.605 68.402 re f 192.124 506.292 214.325 69.122 re s 247.326 574.933 m 247.326 505.812 l S 157.564 596.294 m 185.884 613.334 l S 0.24 w 2 j 0 g 187.444 610.094 m 192.004 617.054 l 183.604 616.574 l 187.444 610.094 l f* 0 j 0.72 w 143.643 561.733 m 178.684 544.212 l S 0.24 w 2 j 176.644 540.972 m 185.044 541.212 l 179.764 547.933 l 176.644 540.972 l f* 0 j 0.72 w 1 g 278.406 519.852 m 280.129 519.852 281.526 518.454 281.526 516.732 c 281.526 515.01 280.129 513.612 278.406 513.612 c 276.684 513.612 275.286 515.01 275.286 516.732 c 275.286 518.454 276.684 519.852 278.406 519.852 c f 278.406 520.212 m 280.327 520.212 281.886 518.653 281.886 516.732 c 281.886 514.811 280.327 513.252 278.406 513.252 c 276.485 513.252 274.926 514.811 274.926 516.732 c 274.926 518.653 276.485 520.212 278.406 520.212 c s 413.529 610.334 47.761 40.801 re f 413.169 609.974 48.481 41.521 re s BT 0 g 420.61 634.815 TD 0 Tc 0 Tw (Single)Tj ET BT 420.61 618.974 TD (Smear)Tj ET 1 g 420.49 513.612 54.721 54.721 re f 420.13 513.252 55.441 55.441 re s BT 0 g 427.57 551.773 TD (Two)Tj ET BT 427.57 535.932 TD (smears)Tj ET BT 427.57 520.092 TD (Per slide)Tj ET 1 g 95.762 572.653 68.402 78.482 re f 95.402 572.293 69.122 79.202 re s BT 0 g 102.602 634.815 TD (Collection)Tj ET BT 102.602 618.974 TD (information)Tj ET BT 102.602 602.894 TD (here in)Tj ET BT 102.602 587.053 TD (pencil)Tj ET 1 g 192.484 335.768 213.605 6 re f 192.124 335.408 214.325 6.72 re s q 48.241 0 0 6.72 192.004 335.528 cm BI /F /LZW /W 50 /H 7 /BPC 4 /CS [ /I /RGB 15 < FFFFFF0000000000000000000000000000000000000000000000000000000000 00000000000000000000000000000000 > ] ID ($ APd. 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DbQ8V-Fb>=CR9$5!GR]/K%s9Ba7D EI Q 0.72 w 313.087 160.684 m 345.546 160.684 371.889 159.178 371.889 157.324 c 371.889 155.469 345.546 153.964 313.087 153.964 c 280.629 153.964 254.286 155.469 254.286 157.324 c 254.286 159.178 280.629 160.684 313.087 160.684 c s 420.13 209.165 m 337.088 170.764 l S 0.24 w 2 j 0 g 335.528 174.484 m 330.248 167.764 l 338.648 167.524 l 335.528 174.484 l f* 0 j 0.72 w 1 g 427.45 188.884 89.042 26.881 re f 427.09 188.524 89.762 27.601 re s BT 0 g 434.29 199.445 TD (Smear of blood)Tj ET 0.24 w 2 j 385.449 263.046 m 385.449 265.926 l 321.847 265.926 l 321.847 263.046 l 385.449 263.046 l f* 0 j 2 j 322.327 270.966 m 309.367 264.486 l 322.327 258.006 l 322.327 270.966 l f* 0 j 0.72 w 1 g 434.41 251.046 102.962 54.481 re f 434.05 250.686 103.682 55.201 re s BT /F2 11.52 Tf 0 g 441.25 289.207 TD (PUSH)Tj /F1 11.52 Tf 30.724 0 TD ( the slide,)Tj ET BT 441.25 273.366 TD (and thus)Tj ET q 441.13 254.646 89.282 47.521 re W n BT /F2 11.52 Tf 441.25 257.286 TD (PULL)Tj /F1 11.52 Tf 30.724 0 TD ( the blood)Tj ET Q 164.524 231.965 m 241.566 174.124 l S 0.24 w 2 j 238.805 171.364 m 247.206 169.924 l 243.366 177.364 l 238.805 171.364 l f* 0 j 0.72 w 1 g 109.443 211.685 68.402 68.402 re f 109.083 211.325 69.122 69.122 re s BT 0 g 116.523 263.526 TD (Keep the)Tj ET BT 116.523 247.686 TD (edge firmly)Tj ET BT 116.523 231.845 TD (against the)Tj ET q 116.403 215.285 54.721 61.441 re W n BT 116.523 213.605 TD (slide)Tj ET Q 1 g 198.965 610.094 41.281 41.521 re f BT 0 g 205.805 635.055 TD (PR)Tj ET BT 205.805 619.214 TD (567)Tj ET 1 g 198.965 513.372 41.281 55.441 re f BT 0 g 205.805 552.253 TD (PR)Tj ET BT 205.805 536.412 TD (568)Tj ET BT 205.805 520.572 TD (568)Tj ET 1 g 382.089 630.494 m 383.811 630.494 385.209 629.097 385.209 627.374 c 385.209 625.652 383.811 624.254 382.089 624.254 c 380.366 624.254 378.969 625.652 378.969 627.374 c 378.969 629.097 380.366 630.494 382.089 630.494 c f 382.089 630.854 m 384.01 630.854 385.569 629.295 385.569 627.374 c 385.569 625.453 384.01 623.894 382.089 623.894 c 380.168 623.894 378.609 625.453 378.609 627.374 c 378.609 629.295 380.168 630.854 382.089 630.854 c s 281.886 527.172 m 281.886 561.493 l S 0.24 w 2 j 0 g 285.607 561.133 m 281.766 568.813 l 277.926 561.133 l 285.607 561.133 l f* 0 j 0.72 w 371.889 630.854 m 316.687 630.854 l S 0.24 w 2 j 317.047 634.815 m 309.367 630.974 l 317.047 627.134 l 317.047 634.815 l f* 0 j 1 g 268.086 637.935 124.323 20.64 re f q 274.806 641.295 110.883 13.92 re W n BT 0 g 274.926 639.855 TD (Direction of smear)Tj ET Q 288.727 513.372 62.161 41.521 re f BT 0 g 295.807 538.332 TD (Direction)Tj ET BT 295.807 522.492 TD (of Smear)Tj ET endstream endobj 14 0 obj 9274 endobj 12 0 obj << /Type /Page /Parent 5 0 R /Resources << /Font << /F1 6 0 R /F2 7 0 R >> /ProcSet 2 0 R >> /Contents 13 0 R >> endobj 16 0 obj << /Length 17 0 R >> stream 0.24 w BT /F1 11.52 Tf 507.732 744.257 TD (3)Tj ET BT 98.762 709.936 TD 0 Tc 0 Tw (5.)Tj /F3 11.52 Tf 8.64 0 TD ( )Tj /F1 11.52 Tf 8.64 0 TD (If doing one smear per slide, the spreader then becomes the next slide to receive a)Tj ET BT 116.043 693.856 TD (smear. Thus, each slide serves two duties, as a spreader, then as a slide to receive a)Tj ET BT 116.043 678.016 TD (smear. If two smears are made per slide, be sure to flip over the spreader to use the)Tj ET BT 116.043 662.175 TD (other edge for the second smear produced. The spreader then is used to receive the)Tj ET BT 116.043 646.095 TD (next two smears. Warning: If there is surplus blood on the spreader, wipe it off)Tj ET BT 116.043 630.254 TD (carefully before flipping it over to make the second smear on the slide.)Tj ET BT 98.762 598.334 TD (6.)Tj /F3 11.52 Tf 8.64 0 TD ( )Tj /F1 11.52 Tf 8.64 0 TD (Photographs are shown in the website.)Tj ET BT 98.762 566.653 TD (7.)Tj /F3 11.52 Tf 8.64 0 TD ( )Tj /F1 11.52 Tf 8.64 0 TD (For blood taken from mammals, a THICK blood film can also be made, but this is not)Tj ET BT 116.043 550.573 TD (possible with blood from birds or reptiles. Only mammals have erythrocytes that)Tj ET BT 116.043 534.732 TD (lack a nucleus. Making a combined thick and think smear for mammal blood is only)Tj ET BT 116.043 518.892 TD (possible if only one smear is made per slide. Make the thin smear starting about 1/3)Tj ET BT 116.043 502.812 TD (from the nonfrosted end of the slide. Then, place another drop of blood at the clear)Tj ET BT 116.043 486.971 TD (end and use the edge of the smearing slide to spread the drop out to about a 1 cm)Tj ET BT 116.043 471.131 TD (circle. The thick smear will take longer to dry. Because the erythrocytes of)Tj ET BT 116.043 455.05 TD (mammals lack a nucleus, thousands of cells can be stacked, and parasites still seen)Tj ET BT 116.043 439.21 TD (\(not for identification, but simply to detect an infected animal\).)Tj ET BT 98.762 407.289 TD (8.)Tj /F3 11.52 Tf 8.64 0 TD ( )Tj /F1 11.52 Tf 8.64 0 TD (Smears should be air-dried, and then dipped into 100% methanol. A coplin jar with a)Tj ET BT 116.043 391.449 TD (screw top is best for this. We use a plastic version, which won\325t break in the field,)Tj ET BT 116.043 375.609 TD (but has a poorly sealing top. Slides can be stored while drying in a small plastic slide)Tj ET BT 116.043 359.528 TD (box \(holds 25 slides\). Then, they are placed, two at a time, back-to-back, into the)Tj ET BT 116.043 343.688 TD (slots in the coplin jar. Thus, ten slides can be dipped at once. Be sure the alcohol)Tj ET BT 116.043 327.848 TD (does not reach the frosted end of the slide. After one minute, the slides are removed)Tj ET BT 116.043 311.767 TD (and placed on end to drain the alcohol. They can then be placed into a plastic slide)Tj ET BT 116.043 295.927 TD (box for complete drying.)Tj ET BT 98.762 264.006 TD (9.)Tj /F3 11.52 Tf 8.64 0 TD ( )Tj /F1 11.52 Tf 8.64 0 TD (In the field, we place the plastic slide box or boxes into a zip-lock bag with silica gel,)Tj ET BT 116.043 248.166 TD (and they are allowed to dry overnight.)Tj ET BT 98.762 216.245 TD (10.)Tj /F3 11.52 Tf 14.4 0 TD ( )Tj /F1 11.52 Tf 2.88 0 TD (To store slides during long field trips, and where many slides are to be made, they can)Tj ET BT 116.043 200.405 TD (be placed back into their original cardboard boxes, with a piece of index card or other)Tj ET BT 116.043 184.564 TD (clean paper between each slide.)Tj ET endstream endobj 17 0 obj 3496 endobj 15 0 obj << /Type /Page /Parent 5 0 R /Resources << /Font << /F1 6 0 R /F3 11 0 R >> /ProcSet 2 0 R >> /Contents 16 0 R >> endobj 19 0 obj << /Length 20 0 R >> stream 0.24 w BT /F1 11.52 Tf 507.732 744.257 TD (4)Tj ET BT /F2 11.52 Tf 98.762 709.936 TD 0 Tc 0 Tw (Field vs. lab preparation of smears \(wild caught animals\))Tj ET BT /F1 11.52 Tf 98.762 678.016 TD (For our work with lizard malaria parasites, we always bring the lizards back into the lab)Tj ET BT 98.762 662.175 TD (in the evening for processing \(even if the \322lab\323 is a hotel room!\), so the smears can be)Tj ET BT 98.762 646.095 TD (made in a somewhat controlled environment. For the work on bird parasites, smears)Tj ET BT 98.762 630.254 TD (must be made at the site of capture \(usually when mist-netting in the early morning, and)Tj ET BT 98.762 614.414 TD (often in web environments\). Very good quality smears are still produced by working on)Tj ET BT 98.762 598.334 TD (the tailgate of a pick-up truck, or on a field table \(a piece of stiff plastic placed on the)Tj ET BT 98.762 582.493 TD (ground\). Smears are kept after dipping in alcohol in a bag with silica gel. Smears made)Tj ET BT 98.762 566.653 TD (in the field in hot and dry climates often are of very poor quality, probably because they)Tj ET BT 98.762 550.573 TD (dry too rapidly. Smears made in the veterinary clinic should be of very high quality)Tj ET BT 98.762 534.732 TD (because of the uniform and clean environmental conditions.)Tj ET BT /F2 11.52 Tf 98.762 502.812 TD (Staining smears)Tj ET BT /F1 11.52 Tf 98.762 471.131 TD (1.)Tj /F3 11.52 Tf 8.64 0 TD ( )Tj /F1 11.52 Tf 8.64 0 TD (First prepare the buffer. The stock buffer should be kept in the refrigerator, but if not)Tj ET BT 116.043 455.05 TD (possible, can be stored at room temperature for several weeks. Make working buffer)Tj ET BT 116.043 439.21 TD (which can be stored at room temperature for a few days. Buffer should be pH 7.0 to)Tj ET BT 116.043 423.37 TD (7.2. Although this is a higher pH than normally used to stain blood cells, the)Tj ET BT 116.043 407.289 TD (parasites will stain darker and be more visible under the microscope.)Tj ET BT 98.762 375.609 TD (2.)Tj /F3 11.52 Tf 8.64 0 TD ( )Tj /F1 11.52 Tf 8.64 0 TD (A high-quality Giemsa should be used. Not all Giemsa stains are equal in quality.)Tj ET BT 116.043 359.528 TD (We place a layer of stain in the bottom of a glass coplin jar \(about 3 mL\), then add)Tj ET BT 116.043 343.688 TD (buffer to a level that will just cover the slides \(except for frosted ends!\) when they)Tj ET BT 116.043 327.848 TD (are in the jar. A little practice will tell the amount of buffer to add. Place the slides,)Tj ET BT 116.043 311.767 TD (back-to-back into the slots of the jar, and stain at room temperature for about 50)Tj ET BT 116.043 295.927 TD (minutes.)Tj ET BT 98.762 264.006 TD (3.)Tj /F3 11.52 Tf 8.64 0 TD ( )Tj /F1 11.52 Tf 8.64 0 TD (Remove slides, rinse by dipping a few times into plain buffer, then stand on end to)Tj ET BT 116.043 248.166 TD (dry. Some workers prefer to run a thin stream of tap water over the slide to remove)Tj ET BT 116.043 232.325 TD (all the remaining stain; we have not found this necessary. Be sure to wash out the)Tj ET BT 116.043 216.245 TD (coplin jars after each use. If not properly washed, stain builds up inside the jar and)Tj ET BT 116.043 200.405 TD (reduces the quality of staining.)Tj ET BT 98.762 168.724 TD (4.)Tj /F3 11.52 Tf 8.64 0 TD ( )Tj /F1 11.52 Tf 8.64 0 TD (There is no need to cover-ship the slides. Immersion oil can be placed directly on the)Tj ET BT 116.043 152.643 TD (smear for observing under 1000x.)Tj ET endstream endobj 20 0 obj 3496 endobj 18 0 obj << /Type /Page /Parent 5 0 R /Resources << /Font << /F1 6 0 R /F2 7 0 R /F3 11 0 R >> /ProcSet 2 0 R >> /Contents 19 0 R >> endobj 22 0 obj << /Length 23 0 R >> stream 0.24 w BT /F1 11.52 Tf 507.732 744.257 TD (5)Tj ET BT /F2 11.52 Tf 98.762 693.856 TD 0 Tc 0 Tw (Preparing staining buffer)Tj ET BT /F1 11.52 Tf 98.762 662.175 TD (Stock buffers \(two\))Tj ET BT 133.323 646.095 TD (The alkaline stock is Sodium phosphate, dibasic anhydrous, N)Tj /F1 6.72 Tf 286.567 -2.4 TD (2)Tj /F1 11.52 Tf 3.36 2.4 TD (HPO)Tj /F1 6.72 Tf 23.041 -2.4 TD (4)Tj /F1 11.52 Tf 3.36 2.4 TD (, Sigma)Tj ET BT 98.762 630.254 TD (Chemical S-0879. Mix 9.5 gm with distilled water to make 1000 mL.)Tj ET BT 133.323 614.414 TD (The acid stock is Potassium phosphate monobasic anhydrous, KH)Tj /F1 6.72 Tf 303.607 -2.4 TD (2)Tj /F1 11.52 Tf 3.36 2.4 TD (PO)Tj /F1 6.72 Tf 14.64 -2.4 TD (4)Tj /F1 11.52 Tf 3.36 2.4 TD (, Sigma)Tj ET BT 98.762 598.334 TD (P5379, mix 9.07 gm with distilled water to make 1000 mL)Tj ET BT 98.762 566.653 TD (Working buffer: Mix 39 mL of acid stock with 61 mL of the alkaline stock, and 900 mL)Tj ET BT 98.762 550.573 TD (of distilled water. Check pH, and adjust to ph 7 or 7.2 by adding the acid buffer stock to)Tj ET BT 98.762 534.732 TD (lower pH or alkaline to raise pH. Just a very few mL should be necessary to reach the)Tj ET BT 98.762 518.892 TD (required pH.)Tj ET BT /F2 11.52 Tf 98.762 486.971 TD (Other supplies)Tj ET BT /F1 11.52 Tf 98.762 455.05 TD (Microscope slides. Good-quality slides seldom will retain any oil from machines used in)Tj ET BT 98.762 439.21 TD (their manufacture, so cleaning should not be required. We use slides with frosted end)Tj ET BT 98.762 423.37 TD (from VWR \(#48311-950\).)Tj ET BT 98.762 391.449 TD (Giemsa. Not all Giemsa stains are equal in quality. We use Baker obtained from VWR)Tj ET BT 98.762 375.609 TD (No. JTM708-1, a 500 mL bottle. This plastic bottle has a pour spout that ALWAYS)Tj ET BT 98.762 359.528 TD (leaks. So, we store the bottle in a plastic bag and always handle the bottle through the)Tj ET BT 98.762 343.688 TD (bag. Giemsa stain will color skin for several days!)Tj ET BT 98.762 311.767 TD (Slide boxes. In the field we use blue plastic slide boxes that hold 25 slides. These are)Tj ET BT 98.762 295.927 TD (obtained from Carolina Biological Supply \(Carolina Blue Boxes, #HT-63-4200\) \). For)Tj ET BT 98.762 280.086 TD (permanent storage, we use wooden boxes from VWR \(#48450-006\).)Tj ET BT 98.762 248.166 TD (Coplin jars. The plastic jar used in the field for dipping into methanol is obtained from)Tj ET BT 98.762 232.325 TD (Carolina \(#HT-74-2155\). Staining jars are available from many sources \(Carolina has)Tj ET BT 98.762 216.245 TD (them #HT-74-2160\). Most of ours were hand-me-downs from retiring faculty over the)Tj ET BT 98.762 200.405 TD (years.)Tj ET BT 98.762 168.724 TD (Silica gel is from Sigma \(S7500\) that we buy in the 1 kg can.)Tj ET BT 98.762 152.643 TD (Zip-lock plastic bags should be the ones used for freezer storage.)Tj ET endstream endobj 23 0 obj 2879 endobj 21 0 obj << /Type /Page /Parent 5 0 R /Resources << /Font << /F1 6 0 R /F2 7 0 R >> /ProcSet 2 0 R >> /Contents 22 0 R >> endobj 6 0 obj << /Type /Font /Subtype /TrueType /Name /F1 /BaseFont /Times-Roman /Encoding /MacRomanEncoding >> endobj 7 0 obj << /Type /Font /Subtype /TrueType /Name /F2 /BaseFont /Times-Bold /Encoding /MacRomanEncoding >> endobj 10 0 obj << /Type /FontDescriptor /FontName /ArialMT /Flags 32800 /FontBBox [ -255 -208 1021 896 ] /MissingWidth 278 /StemV 93 /StemH 93 /ItalicAngle 0 /CapHeight 718 /XHeight 531 /Ascent 896 /Descent -208 /Leading 42 /MaxWidth 1021 /AvgWidth 551 /Style << /Panose <0508020B0600000000000000> >> >> endobj 11 0 obj << /Type /Font /Subtype /TrueType /Name /F3 /BaseFont /ArialMT /FirstChar 0 /LastChar 255 /Widths [ 0 750 750 750 750 750 750 750 0 278 750 750 750 0 750 750 750 750 750 750 750 750 750 750 750 750 750 750 750 0 750 750 278 278 355 556 556 889 667 191 333 333 389 584 278 333 278 278 556 556 556 556 556 556 556 556 556 556 278 278 584 584 584 556 1015 667 667 722 722 667 611 778 722 278 500 667 556 833 722 778 667 778 722 667 611 722 667 944 667 667 611 278 278 278 469 556 333 556 556 500 556 556 278 556 556 222 222 500 222 833 556 556 556 556 333 500 278 556 500 722 500 500 500 334 260 334 584 750 667 667 722 667 722 778 722 556 556 556 556 556 556 500 556 556 556 556 278 278 278 278 556 556 556 556 556 556 556 556 556 556 556 400 556 556 556 350 537 611 737 737 1000 333 333 549 1000 778 713 549 549 549 556 576 494 713 823 549 274 370 365 768 889 611 611 333 584 549 556 549 612 556 556 1000 278 667 667 778 1000 944 556 1000 333 333 222 222 549 494 500 667 167 556 333 333 500 500 556 278 222 333 1000 667 667 667 667 667 278 278 278 278 778 778 750 778 722 722 722 278 333 333 333 333 333 333 333 333 333 333 ] /Encoding /MacRomanEncoding /FontDescriptor 10 0 R >> endobj 2 0 obj [ /PDF /Text /ImageC /ImageI ] endobj 5 0 obj << /Kids [4 0 R 12 0 R 15 0 R 18 0 R 21 0 R ] /Count 5 /Type /Pages /MediaBox [ 0 0 612 792 ] >> endobj 1 0 obj << /Creator (Microsoft Word 98) /CreationDate (D:20050725111313) /Subject () /Title () /Author (jschall) /Producer (Acrobat PDFWriter 4.05 for Power Macintosh) /Keywords () >> endobj 3 0 obj << /Pages 5 0 R /Type /Catalog /DefaultGray 24 0 R /DefaultRGB 25 0 R >> endobj 24 0 obj [/CalGray << /WhitePoint [0.9505 1 1.0891 ] /Gamma 1.8008 >> ] endobj 25 0 obj [/CalRGB << /WhitePoint [0.9505 1 1.0891 ] /Gamma [1.8008 1.8008 1.8008 ] /Matrix [0.3954 0.2208 0.0411 0.4022 0.6391 0.1576 0.1528 0.1405 0.8903 ] >> ] endobj xref 0 26 0000000000 65535 f 0000025678 00000 n 0000025517 00000 n 0000025870 00000 n 0000003649 00000 n 0000025564 00000 n 0000023776 00000 n 0000023889 00000 n 0000000017 00000 n 0000003629 00000 n 0000024001 00000 n 0000024306 00000 n 0000013140 00000 n 0000003790 00000 n 0000013119 00000 n 0000016843 00000 n 0000013271 00000 n 0000016822 00000 n 0000020547 00000 n 0000016975 00000 n 0000020526 00000 n 0000023645 00000 n 0000020690 00000 n 0000023624 00000 n 0000025959 00000 n 0000026039 00000 n trailer << /Size 26 /Root 3 0 R /Info 1 0 R /ID [] >> startxref 26208 %%EOF