lyostat3 freeze drying microscopy
Post on 05-Dec-2014
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Lyostat3Third GenerationFreeze-Drying Microscope
Lyostat3
• Lyostat3 was developed by Biopharma Technology Ltd to provide a rational scientific basis for freeze-drying cycle development. The Lyostat3 combined with BTL’s extensive experience in the freeze-drying industry provides a powerful tool for freeze-drying R&D Scientists
• Biopharma Technology Ltd has used Lyostat3 to analyse over 700 different formulations, from small drug molecules to large complex biomolecules for biopharmaceutical companies worldwide
Freeze-drying microscopy
• Enables real-time observation of the behaviour of your formulation during freeze-drying
• Obtains vital information on the critical temperatures of your product relevant to freeze-temperatures of your product relevant to freeze-drying
• Enables you to develop a safe and efficient freeze-drying cycle specific your product
Benefits of using Lyostat3
• It can help improve old and develop new formulations
• It can provide essential data to develop cycles on a rational scientific basis
• It can save valuable development time and money• Enables rapid determination of collapse, melting and
eutectic event temperatures• Archives data and image capture for future
reference• Only 2µl of sample material is required to determine
the key parameters
Lyostat3 System Components
System Components
• Lyostat3 Freeze-Drying Cryostage– Temperature range -196°C to +125°C– Up to 130°C per min heating/cooling rates– Silver heating block for high thermal conductivity
• Innovative New Imaging Station– Purpose Built– Small benchtop footprint – Higher resolution than standard compound microscopes – Tilting lens and camera for easy loading of samples
System Components
• Temperature Control System– Temperature is controlled by carefully balancing heat
input from a heating element and the precise control of liquid nitrogen through the silver block
– Temperature is measured to 0.01°C by a Pt100 – Temperature is measured to 0.01°C by a Pt100 resistor mounted close to the sample and controlled to 0.1°C even down to pressures as low as 10-3mBar
System Components
• High Performance Digital Firewire Camera• Dedicated Digital Image Capture Software
– Displays real-time temperature and pressure information and gives the user full control heating rate, limit and hold time of up to 100 ramps within the temperature profile.
– Image capture is integrated with temperature programmer data – Image capture is integrated with temperature programmer data capture.
– Images can be loaded into a gallery of either 2,8 or 32 images, for closer inspection.
System Components• Vacuum Pump
• Vacuum Gauge
• Optional Motorised Vacuum Control System– Software-controlled motorized valve controls chamber pressure– Enables close investigation of the effects of pressure on sample
collapsecollapse– Chamber pressure displayed in Torr, microns, mBar and Pa– Pressure plotted with temperature against time and can be
saved for later analysis.
Assay Procedure
• Unscrew lid and side door• Place sample holder on
block and close door• Place drop of silicone oil
on sample block• Place quartz cover slip in
Sample Block
• Place quartz cover slip in the sample holder
• Place spacer ring on quartz cover slip
• Pipette 2µl of sample onto quartz cover slip
• Place a smaller cover slip on top of the spacer ring
Sample Holder
Stage Lid
Quartz cover slip (16 mm dia.)
Glass cover slip (13 mm dia.)
2µl of sample
Objective Lens (usually 10x)Metal Spacer (70µm thick)
Assay ProcedureSample Loading
Drop of silicone oil for low
Temperature Controlled Block
Light Source (from below)
Aperture
dia.)
Sample Holder
Drop of silicone oil for low temperature lubrication
Assay Procedure
• Once sample is loaded onto stage it can be viewed in real-time on the computer screen
• Program your freezing temperature and initiate
Freezing
temperature and initiate the pump with one click
• When your freezing temperature is attained it can be held to ensure complete freezing
• Turn on the vacuum pump to begin drying the sample
Assay ProcedureDrying – Amorphous Product
Frozen Amorphous
Material
When drying, the sublimation
front will move from one side
of the sample to the other. In
front of it will be the frozen
Dried Amorphous
Material
Movement of sublimation Front
front of it will be the frozen
material and behind will be
the dried material
Sublimation Front
The collapse temperature (Tc) is obtained by gradually increasing the temperature until product collapse is observed
Assay ProcedureDrying – Amorphous Product
Frozen Amorphous
Material
Raising the temperature above the
Tc of the sample will cause it to
undergo collapse
A precise T can be found by going
Dried Amorphous
Material
A precise Tc can be found by going
back and viewing the images
recorded during the analytical runCollapsed Material
For crystalline products the same method is applied but a eutectic temperature
(Teu) is obtained instead
Assay ProcedureDrying - Crystalline Product
Sublimation Front
Eutectic melt of material at interface
Frozen Crystalline Material
Dried Crystalline Material
Change in gross morphology of frozen phase to give more liquid
appearance
Increasing Temperature
Crust Formation
Image of a solution showing evidence of crust formation
The black line around the sample
edge is an area of higher solute
concentration that impedes vapour
flow and inhibits drying
When the crust ruptures the sample will
dry from the site of the break. In this
case the temperature had been raised
above the sample Tc and so shows
evidence of collapse
Micro-collapse
Image of a 2% Mannitol + 1% Glucose solution drying under vacuum at -41.8°C, around the Tc of Glucose
Frozen material
Movement of sublimation front
Dried material
Mannitol retains
structure
Glucose collapses
within mannitol structure
Create MovieView Pictures
Data Overview
This file contains all the
information from the analytical
run
Results Data
From here you can view the
pictures recorded in a gallery
or put them together to create
a movie (.avi format)
The data chart shows temperature and pressure information throughout the
procedure
Data OverviewEach point on the purple image line
represents a single image viewed by
selecting it with the cursor
The single image view enables the user to
draw lines and a size bar to measure
Results Data
draw lines and a size bar to measure
between points of interest on the image
(e.g. ice crystal size)
Date, time, vacuum, temperature and ramp
information are displayed under the image
Image Gallery
The gallery view enables
fast sorting of images,
showing how the drying
Gallery Navigation
ZoomImage Number
Results Data
showing how the drying
progresses across a longer
timeframe
For larger images the gallery can be viewed showing less images – 32, 8 and 2
are available
Determination of critical temperatures
‘Trial and Error’ Cycle resulting in loss of product or costly inefficient cycle
New cycle based on actual
parameters
Problem Solved
Lyostat3 provides you with essential information on the critical temperatures of your formulation, enabling you to get the best results
from your product
Biopharma House, Winnall Valley Road, Winchester SO23 0LD, UK
Tel: +44 (0)1962 841092 Web: www.btl-solutions.net
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