圖片	品牌	名稱	貨號	規(guī)格	價格
	Glycotech	Rectangular Flow Chamber Kit	31-010	1 kit	詢價
	Glycotech	Circular Flow Chamber Kit	31-001	1 kit	詢價

7. Dynamic Flow Assay in a Parallel Plate Flow Chamber

John T. Patton~GlycoTech Corporation, Rockville, Maryland 20850

Flow assays allow visualization of cell adhesion under well-defined wall shear stress. The visualization of the different events of cell adhesion can be quantified by selective image acquisition and subsequent image processing. Flow assays are uniquely suited to the investigation of adhesive events which occur very rapidly in a time scale shorter than that of most static adhesion assays. In addition, events subsequent to the initial events can be studied such as cell stabilization and spreading giving some insight into the kinetics of particular cell-cell or cell-substrate adhesive behavior.

Materials:

Microscopy: Inverted-stage microscope configured for phase contrast and fluorescence operation Objective lens (6.3 x, 10 x, 40 x) Stage incubator	Biological: Cell suspension (neutrophils, cancer cells) Cell monolayer or coated substrate in 35 mm dishes Adhesion media (culture media serum-free with 12 mM Hepes) Fibronectin (human plasma) Bovine serum albumin (BSA)
Flow Chamber System: Flow chamber deck with gasket and tube fittings 35 mm tissue culture dishes Harvard syringe pump Glass syringes to fit on pump (3, 5, 20, or 60 cc) Silastic tubing Stopcocks Vacuum pump	Computer/Electronics: Digital image acquisition and processing system Display monitor Video recorder CCD camera with controller Time-date generator Computer with interface to image system Storage device for images

Protocol:

Cell Monolayer Preparation

1. Coat dishes with human fibronectin (FN) by adding 1 ml of 5.5?g/ml FN solution to each dish.

2. Incubate for 30 min. at room temperature.

3. Aspirate solution out of each dish.

4. Add 2 ml of cell suspension (Huvecs, CHO cells) to each dish at seeding density of 0.5-3.0 x 105 cells/ml depending on time required to reach confluence.

5. After 1-5 days, use dishes with confluent monolayer in flow assay. Feed cells every 2-3 days, but usually not with 48 hrs. of flow assay.

Coated Substrate Preparation

1. Outline a coating region (5 mm diameter) for coating substrate in the center of each dish with marking pen.

2. Add 20?l of solution containing substrate at a concentration of 10?g/ml to coating region. Incubate 1 hr.

3. Aspirate off liquid, add 20?l of 1% BSA to block for 1 hr.

4. Aspirate off BSA, add 20?l of inhibitor for 1 hr.

5. Dishes are ready for use in flow assay.

Flow Assay Using Parallel Plate Flow Chamber

1. Turn on stage incubator and warm adhesion media to 37?C 1 hr. prior to beginning flow assay.

2. Assemble flow system apparatus connecting inlet, outlet, and vacuum lines to the flow chamber deck. Fill system with media and remove all air from system.

3. Fill inlet reservoir with cell suspension. For assays using cell monolayers, 106 cells/ml is recommended. For coated substrate assays, use 105 cells/ml.

4. Attach dish to flow chamber deck by holding the deck inverted, place a small bubble of media on flow path area, then place 35 mm dish on the deck. Vacuum will hold dish on deck. Make sure dish was attached with no air bubbles in the flow path.

5. Place assembled chamber on microscope stage.

6. Initiate flow of cells syringe pump connected to outlet flow chamber at a shear stress in the range of 0.1-4.0 dynes/cm2.

7. Allow cells to flow for sufficient time to get an adequate number of cells interacting with the cell monolayer or coated surface. Generally 3-10 min. is used.

8. Begin image acquisition. Collect images at 7-10 locations on the dish. Generally 3 dishes at a given experimental condition gives enough data to show statistical differences between treatments.

9. After images are acquired on all dishes, perform image analysis to quantify the flow assay.

References:

(1) Lawrence, M.B., McIntire, L.V., Eskin, S.K., (1987), Effect of flow on polymorphonuclear leukocyte/ endothelial cell adhesion, Blood 70: 1284-1290.

(2) Patton, J.T., Menter, D.G., Benson, D.M., Nicolson, G.L., McIntire, L.V., (1993), Computerized analysis of tumor cells flowing in a parallel plate chamber to determine their adhesion stabilization lag time, Cell Motility and the Cytoskeleton26: 88-98.

(3) Jones, D.A., Abbassi, O., McIntire, L.V., McEver, R.P., Smith, C.W., (1993), P-selectin mediates neutrophil rolling on histamine-stimulated endothelial cells, Biophysical Journal 65: 1560-1569.

NE-1000程控注射泵,貨促銷

NE-1000 Programmable Single Syringe Pump

技術參數(shù)：

注射器的容量達到60ml 
注射速率可以從0.73uL/hr-2100mL/hr調節(jié) 
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1.有注入和回抽功能 
2.可編程控制，*大41階命令（注射的速率、注射的容量、插入暫停） 
3.一臺電腦可以控制100臺注射泵 
4.注射的精度小于正負1%

New Era系列注射器微量注射泵可用于動物實驗，同時具有注射和回抽功能，可控制流速，可通過程序控制注射流量。相比較其它同類產(chǎn)品，New Era泵提供了更強的功能，然而價格確比其它品牌要低。這讓New Era泵嬴得了大量的客戶?？蛻粝蚱渌耐峦扑]這種微量注射泵。對于需要雙泵的用戶而言，New Era系統(tǒng)的優(yōu)勢是巨大的。它以不高于其它品牌雙通道泵的價格，提供兩臺單通道泵。這兩臺泵可以單獨控制，從事不同的實驗，也可以互聯(lián)，運行相同的參數(shù)或程序。功能和實用性均遠超其它廠家的雙通道泵。

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主要特性:

1. 可單機操作也可電腦控制 (RS-232插頭)

2. 可注射也可回抽 (雙向)

3. 可用1~60cc注射器

4. 可選擇注射流量及流速控制程序，有41種選擇

5. 可將多個泵連接組合成一個系統(tǒng) (雙筒), *大可達100泵;

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NE-1000: 0.73 ml/小時~2100ml/小時

NE-4000: 0.73 ml/小時~2100ml/小時

NE-1600: 0.568 ml/小時~1337ml/小時

NE-1800: 0.568 ml/小時~380 ml/小時

技術參數(shù):

注射器尺寸　　　 1-60ml,or 0.5-5μl微量注射器

注射器數(shù)目　　　　　1，6，8　依據(jù)泵的型號變化

驅動方式　　　　 步進馬達，1/8 to 1/2步進方式

轉軸*大步進數(shù)　 400/s

馬達步進距離　　 0.850μm(1/2step)

馬達/轉軸齒數(shù)　　　 15/28

速度（max/min） 5.1cm/min 0.0042cm/hr

泵流量　　　　　　　*大：1699mL/hr，60ml注射器

　　　　　　　　　　*?。?.73μL/hr，1ml注射器

*大推力　　　　　　35lb *小流速時

　　　　　　　　　　18lb *大流速時

程序數(shù)　　　　　　　41

電源　　　　　　　　220V，50-60Hz

尺寸　　　　　 　22.9×14.6×11.4 cm

重量　　　　　 　1.6kg

 

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