Materials and Reagents

1. Arabidopsis thaliana inflorescence stems
   
2. MS salt mixture (Wako Pure Chemical Industries, catalog number: 392-00591 )
3. 1% (w/v) sucrose
   
4. 0.05% (w/v) MES
   
5. 0.1% (w/v) agar
   
6. Growth media (see Recipes)
   

Equipment

1. Fine tweezers
   
2. Scissors
   
3. Razor blade (Electron Microscopy Sciences, catalog number: 72000 )
   
4. Kimwipes
   
5. Aluminum chamber (custom built) (NSK Ltd.)
   
6. Silicone rubber (thickness: 0.5 mm) (AS ONE Corporation, catalog number: 6-611-01 )
   
7. Round cover glass (diameter: 12 mm) (Matsunami Glass, catalog number: CO12001 )
   
8. CMS system (Figure 1, not commercially available) (NSK Ltd., http://www.nsk.com/)
   CMS is a newly designed compact centrifuge microscope, 30 cm in height and 20 cm in diameter. CMS consists of a direct-drive motor (NSK Ltd., MEGATORQUE MOTOR™, model: M-PS1006KN002 ) and optics including a 50x objective lens with a working distance of 18 mm (SLMPLN 50x, 0.35 NA, OLYMPUS), LED light source (SCHOTT MORITEX Corporation, model: MEBL-CW25 ) and a CCD camera (SENSOR TECHNOLOGY, model: STC172C ).
   
   
   Figure 1. Overview of the CMS system
   
   
9. Windows computer [minimum computer requirements: Windows^® XP or later, Pentium M 778 (1.6 GHz), RAM 1024 MB or higher]
   Note: To install the software below, Windows computers are highly recommended.
   
   

Software

1. MEGATORQUE MOTOR™ controller (EDC megaterm software) (NSK Ltd., http://www.nsk.com/)
   Note: This is free software to control the motor and is available only for Windows.
   
2. Video capture software (COREL, http://www.corel.com/)
   Note: You can use any video capture soft/hardware that converts analog video signal into digital signal and stores the data in a computer.
   
3. G-Track spot-tracking software (G-Angstrom, http://www.g-angstrom.com/eng/products/index.php)
   Note: This is a piece of commercial software to trace fluorescence/bright spots and available only for Windows.
   

Procedure