Leica VT1000 A Vibratome Operating Manual Leica VT1000 A V1.1 English - 01/2010 Always keep this manual with the instrument. Read carefully before working with the instrument.
NOTE The information, numerical data, notes and value judgments contained in this manual represent the current state of scientific knowledge and state-of-the-art technology as we understand it following thorough investigation in this field. We are under no obligation to update the present manual periodically and on an ongoing basis according to the latest technical developments, nor to provide our customers with additional copies, updates etc. of this manual.
Contents 1. Important Notes......................................................................................................................................................... 5 2. Safety......................................................................................................................................................................................6 3. 2.1 2.2 Instrument Components and Specifications...............................................................................
1. Symbols in the text and their meanings Dangers, warnings and cautions appear in a gray box and are marked by a warning triangle. Notes, i.e. important information for the user, are highlighted in gray and marked by the symbol. (5) (Fig.3) Numbers in parentheses refer to item numbers in illustrations or to the illustrations themselves. Qualification of personnel • The Leica VT1000 A may be operated by trained laboratory personnel only.
2. Safety The safety and caution notes in this chapter must be observed at all times. Be sure to read these notes even if you are already familiar with the operation and use of other Leica products. 2.1 Safety notes This Operating Manual contains important instructions and information regarding the operational safety and maintenance of the instrument. The Operating Manual is an important part of the product, and must be read carefully prior to startup and use and must always be kept near the instrument.
2. Safety Warnings – Markings on the instrument itself • Warning labels on the instrument marked with a warning triangle indicate that the correct operating instructions (as defined in this Operating Manual) must be followed when operating or replacing the item marked. Failure to adhere to these instructions may result in an accident, personal injury, damage to the instrument or accessory equipment. • Observe the warning messages on the rear side of the instrument (Fig. 2).
2. Safety Warnings – Operating the instrument • Take care when handling disposable blades and sapphire blades. The cutting edge is extremely sharp and can cause serious injuries! Always wear work safety shoes and safety gloves! • Always clamp the specimen block BEFORE clamping the blade/knife. • If making adjustments or configuring settings in the specimen area, take the blade out of the holder in order to prevent cutting injuries from accidental contact with the blade.
3. 3.1 Instrument Components and Specifications Overview — instrument components LED Light Assembly Magnifier Specimen Bath Vibratome tray Section Thickness Knob Adjustment knobs for the sectioning window Control panel Rear view of the instrument Hazard warning Nameplate Fuses Connection for footswitch Power switch Connection Power cable Drainage valve for liquid from the Vibratome tray Foot switch ON/OFF Leica VT1000 A Instrument feet Fig.
3. Instrument Components and Specifications 3.2 Technical data General Approvals: The instrument-specific approval marks are located on the identification label. Operating temperature range: +10 °C to +35 °C Relative humidity: max. 80% non-condensing Operating temperature range during storage: + 5 °C to +55 °C Storage humidity: < 80 % Microtome Max.
4. 4.1 Setup the Instrument Standard delivery The standard equipment of the Leica VT1000 A is available in 3 variants and includes the following parts: 1. STARTER PACKAGE 1 Specimen Tray (includes V-block adapter).................................... 39053744 1 Specimen Blocks, (3 blocks)............................................................. 39053747 2 Specimen Adhesive........................................................................... 39053986 2 Feather Blades, (20 blades)..................
4. Setup the Instrument 3. PROFESSIONAL GOLD PACKAGE 1 1 1 4 5 1 1 1 1 1 2 1 1 Rotating Stage Assembly.................................................................. 39053760 Foot Pedal............................................................................................ 39053300 Sapphire Knife..................................................................................... 39053237 Specimen Adhesive...........................................................................
4. 4.3 Setup the Instrument Unpacking and setting up the instrument • First check the shipment for external damages upon arrival. • If it is evident that the shipment was damaged during transport, please make a claim to the carrier immediately. • Ensure that the instrument is standing on a work surface that is as free of vibrations as possible. The instrument must be set up the instrument so that the left rear side of the instrument (item 15 in Fig. 7) is easily accessible at any time. 1.
4. Setup the Instrument 4.4 Connection to power supply system 4.4.1 Checking the performance requirements The Vibratome VT1000 A is available in different versions for operation with different supply voltage.
4. Setup the Instrument 4.4.2 Connecting to the power supply All electrical connections are on the left rear side of the instrument. 15 16 The Leica VT1000 A MUST be connected to a grounded power socket. The instrument is supplied with a power cable that is suitable for the local power supply (socket). Only this cable may be used to operate the instrument! Do not use an extension cable! Fig.
4. Setup the Instrument 4.5 Connecting the foot switch The standard scope of delivery of the "Professional" and "Professional Gold" instrument versions include a foot switch (18, Fig. 8) that can be connected to the rear side of the instrument. In SINGLE and AUTO modes, the sectioning process can be started and stopped with the foot switch instead of the DIRECTION switch.
4. 4.6 Setup the Instrument Assembling the "deluxe specimen tray" 24 25 26 Fig. 9 • Fasten the u-frame bracket (24, Fig. 9) for the deluxe specimen bath in the main specimen bath with the corresponding countersunk hexagon screw (26) on the round specimen stage (27). The deluxe specimen bath (22, Fig. 11) is assembled inside the standard black specimen bath (23, Fig. 10). It enables separation of buffer solution and ice preventing dilution of the buffer while sectioning.
4. Setup the Instrument Assembly of the "deluxe specimen tray" (continued) 25 • As shown in Fig. 12, attach the u-frame bracket to the main specimen bath (27, Fig. 10). • Insert the countersunk screw (26) into the bore and tighten it using a size 3 Allen key (25). When tightening, ensure that the fixture (24) is parallel to the front edge of the Vibratome tray (Fig. 12). 26 Fig. 12 Enlarged detail: Knurled screw and clamping piece for fastening the deluxe specimen tray in the fixture.
4. 4.7 Assembling the specimen vice assembly 23 27 Fig. 15 36 34 35 37 Setup the Instrument Always remove the blade BEFORE installing or removing accessories! • The specimen vice assembly (34, Fig. 16) is assembled inside the main specimen bath (23, Fig. 15). • Insert the cheese head screw (35) into the bore of the specimen stage (27) and tighten it using a size 5 Allen key (37, included in the scope of delivery).
4. Setup the Instrument 4.7.1 Inserting the round specimen tray 45 46 The round specimen tray (45) can be installed in place of the specimen block. It enables easy separation of buffer solution and ice that can accumulate in the main specimen bath. Always remove the blade BEFORE installing or removing accessories! Fig. 19 46 To insert it, follow these steps: • First, insert the V-block adapter (46) into the tensioning clamp (46) as shown in Fig. 20.
4. Setup the Instrument 4.7.2 Assembling the rotating stage assembly 40 • Unscrew the knurled screw (38) completely. You can now take the specimen clamp (40) off of the base plate (39) (Fig. 23). 38 39 Fig. 23 45 38 The rotating stage assembly allows for simple position correction of the specimen surface when the specimen is glued in place. • To do so, first remove the specimen vice assembly (34, Fig. 16) by following the reverse order of steps as described in Chap. 4.7.
4. Setup the Instrument 4.8 Assembling the magnifier The standard scope of delivery of the instrument includes a 2x magnifier that can be assembled on the lamp carrier. 11 11 • Move the lamp carrier (10, in Fig. 27) on the instrument out by pulling it forwards. • The bottom of the lamp carrier has two notches (12), one each on the left and right. • Install the magnifier (13) so that the two pins (11) on the left and right fit into the corresponding groove.
5. 5.1 Operation Operating elements and their functions 5.1.1 The control panel DIRECTIONAL SWITCH Direction of cutting head PAUSE Switch Rotary knob AMPLITUDE Rotary knob SPEED Display Height adjustment of Section thickness Number of the specimen stage sections 5.1.2 The operating elements Light switch Sectioning switch Automatic/Single/ Manual Fig. 28 PAUSE switch With this switch, you can temporarily stop the feed movement of the blade at any place.
5. Operation The operating elements (continued) DIRECTION switch Fig. 30 Switch position: Upwards --> forwards Downwards --> backwards Using this rotary knob, you can change the relative speed of the cutting head (feed movement) if the DIRECTION switch is in the FWD/ AUTO REV position. • In the zero position (‘0‘), the blade does not move; in the ‘10‘ position, it is moved at maximum speed (2 mm/s). • For the positions in between, the speed is changed proportionally. Rotary knob SPEED Fig.
5. Operation The operating elements (continued) AMPLITUDE rotary knob Fig. 32 This knob controls the relative amplitude of the lateral excursion of the sectioning blade vibratory movements. • In the zero position (‘0‘) or during the retraction movement of the cutting head, the blade does not vibrate. • The maximum amplitude (2.0 mm) is reached when the rotary knob is in the ‘10‘ position. At intermediate settings, the amplitude will vary proportionally.
5. Operation The operating elements (continued) THICKNESS switch 53 The THICKNESS switch defines the desired section thickness for automatic mode in microns. • The parameter selected for this value is indicated in the LED display above the switch, the displayed values are specified in µm. The LED (53) in the switch is then illuminated. Switch position: Upwards --> Values for the section thickness are increased. Downwards --> Values for the section thickness are decreased. Fig.
5. Operation The operating elements (continued) MODE switch Fig. 38 The automatic switch also serves as the reset button in automatic mode. If you want to reset the instrument in serial sectioning mode to the default parameters for sectioning operations and section thicknesses, the counters can be reset using the automatic switch. LIGHT switch Fig. 39 Leica VT1000 A The MODE switch serves to toggle between the manual, single and automatic modes.
5. 5.2 Operation Adjustment options on the instrument 5.2.1 Adjusting the blade presentation angle The presentation angle of the sectioning blade relative to the plane of the section is adjustable to suit operating conditions and specimen types. The adjustment can be made by rotating the blade holder (50) on the mounting bar. • The accessories include a blade angle indicator (46) for this purpose. 46 47 Fig. 40 Enlarged detail: Scale of the blade angle indicator and indicator on the blade holder.
5. 5.2.2 Adjusting the specimen and blade Operation Positioning the specimen in the specimen holder You can adjust the lateral position of the specimen in order to attain an approximate centering of the specimen in the specimen tray. Since one of the jaws of the specimen holder is stationary, the use of specimen mounting blocks of varying widths will allow for adjustable specimen mounting. Fig. 42 Adjust the tilt of the specimen holder.
5. Operation Adjusting the tilt of the specimen If the specimen is still not completely horizontal, you can tilt the specimen along another axis (from side to side). To do so, tilt the specimen mounting block slightly when it is clamped in the jaws of the specimen holder. Fig. 44 Complete control of the specimen over all three axes is possible with the rotating stage assembly (see Chap. 7.2, Optional Accessories). Moving the blade holder relative to the specimen In some cases, e.g.
5. Operation 5.2.3 Adjustable sectioning window The VT1000 A features an adjustable sectioning window that you can use you speed up the process of making series sections. Fig. 46 This "window" is the distance the blade travels from a resting retracted position to the front of the bath when it automatically ends the forward stroke and reverses. This corresponds to distance "A" in Fig. 46.
5. Operation Adjustable sectioning window (continued) To make this adjustment, the instrument should be in manual mode. To adjust the front and rear edge of the sectioning window, you have to move the internal limit switch using two rotary knobs (57), which are on the side of the top plastic cover of the Vibratome. The two rotary knobs can be adjusted independently of one another. 57.2 57.1 Fig. 47 • First, adjust the front limit switch that is on the front side of the instrument.
5. 5.2.4 Adjusting the specimen height 27 58 60 59 Fig. 48 Operation The thickness of sections generated is controlled by the raising of the specimen incrementally. Since the sectioning blade remains stationary along the vertical axis, the increment by which the specimen is raised correlates to the section thickness. The height of the specimen is adjusted using a micrometer feed system that is located below the specimen tray.
5. Operation 5.3 Sectioning The Leica VT1000 A offers a wide variety of options for sectioning fixed and unfixed tissue samples of plant or animal origin. It employs a vibrating blade principle, which allows sectioning without freezing or embedding. This makes it possible to prevent the unwanted effects that occur during freezing or embedding, such as the formation of artifacts, change of the morphology, the impairment of enzyme activities and other harmful processes.
Sectioning (continued) When the instrument is switched on for the first time, the blade is automatically moved back in operating state "B" and stopped in state "C". Each sectioning cycle needs only to be initiated by the operator. Once state "A" is activated, the sectioning blade will advance at the operator-selected forward speed and amplitude until it reaches the forward end-of-travel. The blade then switches into operating state "B" automatically until state "C" is reached.
5. Operation 5.3.1 General instructions As a result of the great variety of specimen types, sizes, shapes, states and preparations, etc. that can be encountered while sectioning with the VT1000 A, it is not possible to provide specific recommendations for parameter settings. The optimal settings for the individual applications can be determined only empirically using specimens that are no longer needed and are as similar as possible to the actual specimens to be sectioned.
5. Operation Direct mounting 5.3.2 Inserting the specimens Specimens rigid enough to be held firmly in the specimen holder without damage may be clamped directly. Keep in mind that excessive clamping forces on the jaws of the specimen holder can cause tension in the specimen. Softer specimens that do not have enough strength on their own (such as a leaf) can be placed between carrier strips or two layers of soft material, e.g. balsa wood or low melt agarose, then inserted into the specimen holder.
5. Operation Adhesive Mounting (continued) Note that in all cases requiring adhesive mounting, thickness of the specimen should be kept as small as practical to maximize rigidity. Before sectioning with the VT1000 A, create a uniform, level surface along the cutting edge of the tissue; this will reduce the time needed for cutting the specimen to size using the blade. Be very careful when using the adhesive, as it very easily sticks to human skin.
5. Operation Adhesive Mounting (continued) Specimens small enough to fit on a surface of a specimen mounting block can be affixed directly. The specimen mounting block is then clamped into the jaws of the specimen holder for sectioning. If the specimen requires a larger surface, you can use specimen tray (39053744) or the deluxe specimen bath (39053745). Fig.
5. Operation Inserting the blade Be careful when handling the disposable blades or sapphire knife. The cutting edge is extremely sharp and can cause serious injuries. Before inserting a blade, the specimen to be section should be attached to the specimen plate (clamped into place or affixed) to prevent injuries. 55 54 53 Fig. 52 55 The blade is held in place by a spring clip (53) in the blade holder that presses against the bottom of the blade holder.
5. 5.4 Operation Specimen preparation Prior to sectioning, gross trimming of the specimen top surface with the sectioning blade to achieve flatness is required. After the specimen and mounting block have been clamped into the specimen holder in the desired orientation relative to the blade feed d irection, the top surface of the specimen should be kept approximately horizontal.
5. 5.5 Operation Preparing the specimen bath The purpose of the specimen bath is four-fold: to lubricate the blade during the sectioning operation, to prevent the specimen from heating up or drying out, to increase or maintain desired features of the specimen and to allow effortless removal of the fragile sections. Any fluid, compatible with polyproylene and nitrile rubber, that would best preserve or maintain the phenomenon or process for which the specimen is being studied may be used.
5. 5.6 Operation Conventional sectioning After the specimen has been trimmed to create a flat specimen surface, the preparations for actual sectioning can be carried out. The procedure consists of generating a few sections at the final parameter settings and section thickness. This is intended to compensate for the effect of lifting the specimen described in the section entitled "General instructions".
5. Operation 5.7 Removing the sections Sections generated may be retrieved from the specimen bath by various means. For microscopic applications, the section may be delivered to a glass slide by aspirating the section with a small glass dropper if doing so does not damage the specimen. In some cases, using a fine sable brush provides more satisfactory results, since the risk of section damage is reduced.
6. 6.1 Cleaning and Maintenance Cleaning the instrument • Always remove the blade before detaching the blade holder from the instrument! • Always keep the sapphire blades in the box when not in use! • Never place used blades on the laboratory table - dispose of them safely! • When using detergents, comply with the safety instructions of the manufacturer and observe the laboratory regulations valid in the country of use.
6. Cleaning and Maintenance Specimen bath area After sectioning is completed, the specimen bath, specimen holder and sectioning blade holder should be cleansed of any bath solution residue. Any water-soluble solvent may be used unless it corrodes parts made of polypropylene or nitrile rubber. The final flushes should be with clean water. This procedure is intended to premature corrosion of metal components in this area, buildup of substances or contamination of subsequent baths.
6. 6.2 Cleaning and Maintenance Maintenance instructions Only authorized, qualified Leica service personnel may access the internal components of the instrument for service or repair! When used normally, the Vibratome VT1000 A requires only standard maintenance. To ensure trouble-free operation of the instrument over a long period of time, the following is recommended by Leica: • Thoroughly clean the instrument on a daily basis. • From time to time, oil the specimen holder (see Fig.
6. Cleaning and Maintenance 6.3 Replacing components 6.3.1 Fuse If the instrument fails to function completely, first check for power availability at the power outlet. The instrument fuse(s) located at the rear of the instrument should be checked next. Before removing the fuse, pull the POWER PLUG OUT OF THE SOCKET. The fuse and its holder are removed by lifting the tab in the fuse drawer that is located directly above the plug and pulling the drawer out of the socket.
7. Optional Accessories 7.1 Ordering information Specimen mounting block, 3 per package........................................... 39053747 Specimen adhesive.................................................................................. 39053986 LED light..................................................................................................... 39053226 Feather Blades, 10 per package............................................................ 39053234 Specimen Tray.............................
