This presentation will describe two microfluidic techniques that my research group has developed for single-cell manipulation and analysis. The first is called Digital microfluidic Isolation of Single Cells for -Omics (DISCO), a platform that allows users to select particular cells of interest from a limited initial sample size and connects single-cell sequencing data to their immunofluorescence-based phenotypes. Specifically, DISCO combines digital microfluidics, laser cell lysis, and artificial intelligence-driven image processing to collect the contents of single cells from heterogeneous populations, followed by analysis of single-cell genomes and transcriptomes by next-generation sequencing, and proteomes by nanoflow liquid chromatography and tandem mass spectrometry. The second is called the optoelectronic microrobot, a system in which optically induced dielectrophoresis is used to control sub-mm particles or "microrobots" that can take nearly any desirable shape or form, and can be programmed to carry out sophisticated, multiaxis operations to manipulate suspended cells. We have used the system for single-cell isolation, clonal expansion, RNA sequencing, manipulation within enclosed channels, controlling cell–cell interactions, and isolating precious microtissues from heterogeneous mixtures. The two systems join a growing list of innovative techniques developed by the microfluidic community that are pushing the boundaries of what is possible for single-cell manipulation and analysis.

Learning Objectives: 

1. Define Digital Microfluidics (DMF)

2. Define Digital microfluidic Isolation of Single Cells for -Omics (DISCO)

3. Define the Optoelectronic Microrobot
 

Explore the journey of the sample through an automated LC/MS workflow from sample receipt to the reporting of results back into a LIMS. The advantages of automation are discussed, along with automation options for the clinical LC/MS workflow based on different laboratory needs. Sample tracking is a key part of any workflow and have been considered in all automated options shown, including other important requirements to ease the extraction and analysis process for the analyst.

Learning Objectives:

1. Why automation is important for LC/MS workflows

2. What can be automated in LC/MS workflows

3. How automation can improve analytical performance and productivity

With the ever-increasing demand for antibody and protein-based therapeutics, a flexible purification platform that can handle low to high sample volumes and expression levels is critical for screening. Protein purification using traditional chromatography is limited by throughput and requires time-consuming, labor-intensive sample preparation (centrifugation and filtration) processes to avoid column clogging and to achieve volumes that are amenable to this platform. Magnetic beads-based purification permits the incubation of the beads directly into cell culture (for secreted proteins) or crude lysates regardless of sample volume. This provides a simplified approach to direct target capture while eliminating much of the sample preparation steps and potentially improving the quality of the purified product. The tools and their application to simplify and significantly augment protein purification and screening cost-effectively will be described. 

Learning Objectives:

1. Learning about purifying protein and antibodies in just 45 to 50 minutes

2. Eluting with minimum 0.4 ml buffer in a 50 ml tube

3. Complete process of washing and elution done automatically

Single cell secretion assays have the potential to dramatically accelerate antibody discovery workflows. This talk will begin with a brief overview of common antibody discovery methods, concepts in single cell partitioning, and some common single cell assays. It will then discuss how bead-based clustering, lensless imaging, and computer vision can be used to perform single cell secretion screening.

Learning Objectives:

1. Understand the antibody discovery protocols in pharmaceutical research

2. Understand the importance of single cell secretion screening protocols

3. Describe lensless imaging and computer vision can be used to perform a chemical assay

4. Describe how Bioelectronica Hypercell performs single cell sorting

Microfluidic technologies have opened up new possibilities for ultra-low volume, and ultrafast assays, compared to bulk-scale counterparts. We have been focused on developing new tools for droplet microfluidics that automate procedures like reagent addition and removal, solid phase extraction, and droplet sorting that can act as plug-and-play components to achieve multi-step analytical processes. Recent advances in droplet microfluidics from our lab will be covered in this talk, including both fundamental device developments and applications in miniaturized assays for protein activity and epigenetic screening, among others.

Learning Objectives:

1. Understand fundamentals of microfluidics and applications for high throughput (bio)chemistry

2. Identify enabling capabilities of droplet microfluidics for compartmentalized profiling.

3. Explain various applications of droplet microfluidics for epigenetic profiling, cell sorting, and high throughput biotechnology.

Analytical Method Validation work is based on a variety of guidelines and requirements hence being time-consuming and stressful. Reducing this work and improving efficiency while ensuring the highest level of data integrity is welcome by both QA and laboratory – but how can this be achieved? Doing method validation is much more than just calculating results – it’s setting up your testing workload, both simple and flexible, with final documentation matching customer expectations as well as regulatory and industry requirements. The key for an optimized workflow is predefining all aspects of typical method validation work by means of powerful software workflow templates that reduce new projects to a matter of a few clicks. Such templates follow your SOPs and structure your tasks to optimize efficiency and compliance. Communication with your laboratory equipment (CDS, LIMS, ECM, ERP, DMS) should be automatic and easy with manual intervention limited to a minimum. While templates define routine work, it is critical to allow flexibility for each new project – additional calculations, graphics and report extensions should be added easily. Since your data is worth being protected and comprehensible all this must be accompanied by a maximum level of data integrity, which means strict access control, full traceability, version history and full 21 CFR Part 11 compliance. This presentation will provide guidance and show optimized software workflows and device integration saving at least 70% of your time compared to performing these tasks manually.

Learning Objectives:

1. Analytical method validation – requirements and data integrity

2. Use efficient template-based workflows to save your time

3. Integrate your lab devices seamlessly into your method validation workflow

For the past years, my laboratory has been working on microfluidic devices to enable the analysis of single cells. Our technology of choice has been droplet microfluidics that encapsulates samples in small (pL-nL range) independent reactors that can be generated and manipulated at rates of 1000’s per second. However, such high throughput loses relevance when analyzing small size samples that contain 100’s to a few 1000’s cells. In addition, there is still a discrepancy between the single-cell manipulation throughput and the capacity of sequencing platforms. To address this unexplored territory, we decided to trade the throughput of droplet microfluidics for higher precision in single cell manipulation and designed a new strategy called trapping and encapsulation. In this approach, we trap almost all the cells (>99%) we inject in our device before encapsulating them. In practice, we came up with different methods to perform the encapsulation. This led us to one particular mode, the capillary-mode, that allows us not only to encapsulate single particles but also to perform controlled and deterministic particle pairs (cells, beads or cell-bead), which has been a long-standing limitation due to the Poisson’s statistics nature of particle encapsulation. Finally, this capillary mode opened new opportunities to perform sample partitioning in plastic material that can be readily manufactured and deployed.

Learning Objectives:

1. Identify various challenges in designing microfluidic devices

2. Explain physical principles harnessed in designing microfluidic devices

3. Review fundamental limitations of current approaches for single particle encapsulation and microfluidic manufacturing

Clinical sample management, particularly for multi-centred trials, has been a challenge for every trial leader for many years and the advent of ever more complicated biomarker sampling has increased the burden further. Aside from the ethical responsibility to look after every patient’s samples, in order to comply with GLPC guidelines, Sponsors have to demonstrate full traceability from the point of sample collection, during transportation, at the analysis lab until finally being transferred to long term storage or destroyed.  Every individual aliquot must be reconciled with the Trial Master File, which typically amounts to numbers exceeding tens of thousands.  Manual process, therefore, leave the door open for errors. This presentation describes how the integration of a LIMS system can simplify your clinical sample workflows and ensure GLPC compliance.  The pitfalls are explored before examining each stage of the sample lifecycle with examples of how LIMS can automate and provide robust processes. The author pulls on a 30-year career in the clinical trial industry to present this overview.

Learning Objectives:

1. Understand the regulatory requirements for clinical sample tracking.

2. Identify the pitfalls of sample tracking during the sample life cycle.

3. Understand how LIMS can alleviate the pitfalls and ensure regulatory compliance.

Arguably data integrity is important, very important. It is clear that with bad data, poor decisions are made, in some cases you might as well have not have any data upon which to base your decisions. So, poor testing along with poor quality data can not only cost companies and governments millions, if not billions of dollars, more importantly it can also result in significant loss of life. During the recent COVID pandemic the need for high quality data, management and control has become even more apparent. We will review the events of the initial attempt by the US government to generate and distribute COVID test kits to health departments through all states. We will examine rejected sampling kits by the Canadian government due to mold contamination and vaccine development challenges by pharmaceutical companies all relating to data integrity. Clinical and analytical testing laboratory data along with meta data is being generated at amazing rates, everything from the specimen demographics, supplies, reagents, specimens tracking, temperature readings of refrigerators, freezers and incubators to ensure that reagents, controls, supplies, and samples are stored at the appropriate temperatures, to data electronically imported from instrumentation (which also generate significant volumes of raw data), result data, dilutions, unit conversions as well as quality control data that all needs to be managed, controlled and maintained. There is a shortage of classically trained microbiologists and professionals that have an aptitude for analytical laboratory work, with a pandemic there is a rapid increase in laboratory testing which puts additional strain on health departments and clinical testing laboratories. This presentation will examine the importance of data integrity, along with the role of LIMS and other tools that can be used to help analytical laboratory professionals to produce, analyze, control, communicate and maintain high quality data. We will look at recent events of failures to generate high quality data and the human and financial impacts during the COVID pandemic.

Learning Objectives:

1. Understand the role that a LIMS plays in defining and maintaining data integrity and why it is so critical.

2. Learn how the impacts of poor data integrity can extend beyond the laboratory environment or an audit finding, to grave human and financial loss.

3. Become familiarized with the role that automation plays in LIMS along with data integrity.

Traditional hybridoma and phage display methods have failed to yield therapeutic antibodies against difficult targets like most GPCRs and ion channels. This presentation will introduce Berkeley Lights’ new Opto™ Plasma B Discovery 4.0 workflow that enables recovery of 1000s of hits by screening up to 100,000 plasma B cells, down-selection of lead candidates by functional screening, and sequencing and re-expression of >1000 functionally-characterized antibodies … all in just 1 week. By maximizing the diversity of antibodies through direct functional profiling of plasma B cells, the Opto Plasma B Discovery 4.0 workflow will allow users to tackle even the most challenging targets.

Learning Objectives:

1. Identify the limitations on hybridoma approaches

2. Understand the unique advantages of the Berkeley Lights Platform

3. Explore current data and case studies behind accelerated discovery against difficult targets

Droplet generators are at the heart of many microfluidic devices developed for life science applications but are difficult to tailor to each specific application. The high fabrication costs, complex fluid dynamics, and incomplete understanding of multi-phase flows make engineering droplet-based platforms an iterative and resource-intensive process. Large-scale experimental datasets and machine learning algorithms are employed to enable performance prediction and design automation of flow-focusing devices with high accuracy. The developed capabilities are captured in a software tool that converts high-level performance specifications to a device that delivers the desired droplet diameter and generation rate. This tool effectively eliminates the need for resource-intensive design iterations to achieve functional droplet generators.

Learning Objectives:

1. Establish machine learning enables accurate performance prediction of flow-focusing droplet generators

2. Establish machine learning enables accurate design automation of flow-focusing droplet generators

3. Introduce an open-source online software for flow-focusing droplet generators, called DAFD

Regulatory agencies across the world are responsible for enforcing regulations for different industries. Industries must adhere to these regulations to assure the quality of their products and services. Getting accredited is expensive, time-consuming, as well as challenging as regulations keep changing. However, ignoring regulatory compliance could be a risk to the quality, public safety, and may lead to heavy penalties. Regulatory requirements may vary from one industry or laboratory to another and from one state or country to another. Some common regulatory compliance and best practices are ISO/IEC 17025:2017, GLP, HIPAA, EU GDPR, 21 CFR Part 11, CLIA, ISO 15189:2012, ISO 20387, GCP, GLP, FSMA, HACCP, and ISBER Best Practices.

Role of a LIMS in Following Regulatory Compliance & Best Practices

A LIMS can help laboratories across different industries, such as clinical, food & beverage, biobanks, to comply with the regulatory requirements and follow best practices. A LIMS helps laboratories in the following ways:

1. Enables following standards and best practices prescribed for an industry.

2. Manages all internal and external documents and tracks their revision history.

3. Automates and streamlines QA/QC processes and ensures traceability through the sample life cycle.

4. Enforces laboratory staff to follow Standard Operating Procedures (SOPs) to minimize procedural errors.

5. Supports validation and approval of multiple test results before releasing the test reports.

6. Maintains staff training records so that analytical tasks could be assigned to the competent staff to maintain the quality of test results

7. Helps schedule equipment calibration, records calibration and maintenance data, and prevents the use of outdated instruments.

8. Generates custom Certificates of Analysis (CoAs) as per reporting standards.

9. Enables maintaining a Chain of Custody (CoC) and a verifiable audit trail.

Learning Objectives:

1. An overview of regulatory compliance and best practices for laboratories across different industries.

2. How a LIMS helps laboratories comply with the regulatory requirements and follow best practices to stay ahead of the curve.

The pandemic sent pharmaceutical and medical device industries scrambling to continue operations under social distancing guidelines. Businesses sent their employees to work from home, yet somehow the work needed to continue, including validation of facilities, utilities and equipment. Learn the regulatory requirements governing pharmaceutical and medical device industries, the fundamentals of validation:  installation qualification, operational qualification, and performance qualification, and ways of leveraging technology solutions to accelerate your validation program.

Learning Objectives: 

1. Understand the regulatory requirements for validation as it relates to equipment

2. Learn the components of installation qualification, operational qualification, and performance qualification

3. Be able to leverage technology to perform validations remotely.