Traditionally, imaging workflows required significant manual interaction and time investment from the user. The addition of automation has decreased user time commitment and increased data output. With the increased data output, there is now the need to effectively analyze the output and turn data into valuable insights. Miltenyi Biotec imaging solutions integrate many automation features into the imaging pipeline to increase the efficiency of the process and accelerate research discoveries.

Learning Objectives:

1. Understand how automation in image acquisition can increase productivity in imaging projects.

2. Explore automation in image analysis to translate biological observation to quantification.

Microfluidic Tech has served millions of lives through the Biotech industry, whether in life science research or clinical diagnostics. Most of these technologies are spawned at academic and research institutions, taking multiple years to materialize outside the lab. While the publication of initial proof of concepts in the lab can be rewarding, society truly benefits when these technologies are released to the market as reliable and easy-to-use products. But this pivot does not happen overnight. Successful commercialization of tech requires the tremendous efforts of an entrepreneurial team skilled in both technology and business development. This process begins with formulating a business plan to secure funding and resources for product development and business growth through building awareness and sales. To make matters more complicated, the commercialization of clinical products takes longer to fruition than research market products because of regulatory constraints. Clinical devices need to be manufactured under a rigorous quality system, and companies are required to submit data to Government agencies showing the safety and efficacy of their products. Even if executed flawlessly, this entire process could take 5-10 years and tens to hundreds of millions of dollars of investment, a daunting endeavor for aspiring entrepreneurs. In this talk, I will share some of the lessons I learned from my two startups, HandyLab and Celsee, both with successful exits, so that it may inspire some of you to take the plunge to benefit so many others.

Learning Objectives:

1.    Discuss how to develop microfluidic technologies into products for life sciences or clinical diagnostics.

2.    Explain how to design milestones for a new product development.

3.    List what the key lessons are for aspiring entrepreneurs for increasing the odds of success.

As regulatory requirements continue to evolve, and instrumentation becomes more sophisticated, laboratories find themselves managing increasing amounts of data, a comprehensive data management plan, including LIMS (Laboratory Information Management System) and automation are critical. Given the vast amounts of data and information that must be collected, managed and maintained, a LIMS is no longer a luxury but a necessity. Leveraging LIMS and automation, no only ensures valuable resource utilization, as knowledge workers should not be copying and pasting data or transcribing data from one system to another. Laboratory automation with the LIMS includes positive ID (barcoding), instrument integration, sensors, CDS (Chromatography Data Systems) and enterprise integration (SAP, CRMs, Statistical, validation, accounting and other software).  Laboratories must manage raw data as well as meta data, along with audit trails of all the data handling systems, LIMS, instrument data management systems, and chromatography data systems. As the final product of the laboratory is data, it is critical that the data be accurate and precise. We will discuss the meaning of data integrity and what constitutes good data, throughout the data life cycle along with understanding the ALCOA+ acronym.  Laboratories must be following electronic records, electronic signatures as well as 21 CFR Part 11, predicate rules, cGMP and GxP. Additional compliance requirements include technical and procedural controls, critical in Pharmaceutical and Medical Devices, along with Standard Operating Procedures, IQ (Installation Qualification), OQ (Operational Qualification) and PQ (Performance Qualification) as part of the Quality Management System. When there is a failure in the laboratories Quality Management System, which encompasses the elements that are defined above, an FDA enforcement action could result in the form of a warning letter, seizure, injunction or criminal prosecution. 

Learning Objectives:

1. Discuss Data Integrity. 

2. Explain the acronym ALCOA+ and the role that LIMS and laboratory automation play in facilitating the ALCOA+ principles. 

3. Discuss the important role that LIMS and Laboratory Automation software play in ensuring Data Integrity, via data import, tracking, audit trails, data management, automated instrument integration, automated reporting, importing sensor data from incubators, fridges and freezers, traceability and more.

A biobank is a laboratory facility that accepts, processes, stores, and distributes samples and data, for use by research institutions. The role is to store these collections and to keep biospecimens in a safe physical location while maintaining quality so that researchers can access them, and the data associated with these biospecimens. Biobanks play an essential role in biomedical research.
 
ISO recently published a new standard specifically for biobanks, ISO 20387:2018, in response to the growing role of biobanks within the life science industry. This standard covers requirements for all aspects of managing biobanks, including structural, personnel, facilities, processes, and equipment. The benefits of a biobank being accredited make any effort worthwhile as it promotes confidence, assures reliability, reduces costs, and boosts efficiency. Furthermore, ISO 20387 promotes transnational harmonization of biobanking practices and fosters worldwide collaboration to further advance research in healthcare and personalized medicine. 
 
Biobanks can take advantage of informatics tools, such as a biobanking LIMS, to securely manage data, safeguard the personal health information of patients, assign role-based rights to staff for several activities, and define quality management systems to make available samples that are fit for use in research. This, in turn, enables biobanks to meet the requirements of ISO 20387 and meet the needs of the research community. 

A LIMS in the cloud offers several advantages for managing biobank data. It leverages the power of the cloud for resource utilization, storage flexibility, security, and scaling benefits, like no upfront investment is needed for hardware such as servers or networks.

Learning Objectives:

1. Discuss the importance of ISO 20387:2018 accreditation.

2. Discuss general, structural, resource, process, and quality management system requirements of ISO 20387 compliance.

3. Discuss how biobanks can comply with ISO 20387 to demonstrate operational competency and maintain high-quality, annotated samples fit for use in research.

The global pandemic has increased focus and scrutiny on molecular diagnostic assay development, resulting in a need for assays that provide quick results while delivering expected performance and quality. The need for these types of assays in infectious disease diagnostics has become apparent on a wide scale, but molecular diagnostic testing is also poised to grow in other areas such as genetic testing, prenatal testing, oncology, and pharmacogenomics over the next several years.

Growth in the amount of testing being done in these areas brings additional challenges due to a number of issues such as various sample types, presence of inhibitors, and low sample amounts—all of which contribute to potential poor results due to less than ideal amplification reactions.

Learning Objectives:

1. Demonstrate how lyo-ready enzymes can overcome many of these issues and provide a viable solution for assay development.

2. Discuss advantages of lyophilization compared to traditional methods.

3. Explain why the quality and manufacturing considerations are crucial when selecting a commercialization partner.

Cancers are genetic diseases driven by recurrent sets of somatic mutations. Different mutations associate statistically with distinct disease risks and can therefore be useful prognostic markers. But, for the majority the underlying biological mechanisms are unknown. Uncovering the physiological consequences of these mutations is important as this could identify novel targets for treatment. However, since most cancers are polyclonal tissues, multiple distinct mutations contribute to the tumor behavior and linking genotype to phenotype is complicated through the convolution of these clones. Methods with single cell resolution can address this issue and  through simultaneous measurements of genotype and phenotype cause and effect can be linked.

In my talk I present DAb-seq, our new droplet microfluidic sequencing method that profiles single-cell genotypes through multiplexed PCR panels and immunophenotypes through DNA tagged antibody panels. I highlight the unique capabilities of DAb-seq with case studies on leukemia and samples from patients with HIV. As DAb-seq is purposely designed modular we are actively extending the approach to access even richer information in the future.

Learning Objectives:

1. Explain the inherent advantages of high throughput single cell methods.

2. Discuss why joint measurements of genotype and phenotype important to understand genetic diseases.

3. Discuss why genotype phenotype mapping can be useful in the study of infectious diseases.

High-throughput screening (HTS) is crucial to generate hits and leads in drug discovery efforts. Most high-throughput biochemical assays are based on optical detection or radioactivity with high sensitivity and a detection speed of sub-second per well. These assays usually require labeling of substrates/products or coupling reactions. False positives for optical assays and material handling for radioactivity assays can lead to detection artifacts, increase cost, and delay progress in a drug discovery program. Considering the benefits of specificity that MS-based HTS assays offer, there is increased interest in recent years to develop alternative strategies to improve throughput using robust chromatography-free MS methods. Examples of such implementations include the use of matrix-assisted laser desorption ionization (MALDI), solid phase extraction (SPE)-based RapidFire system and ambient ionization techniques such as acoustic droplet ejection and acoustic mist ionization (AMI).

Infrared matrix-assisted laser desorption electrospray ionization (IR-MALDESI) is a hybrid ambient ionization method that combines the use of laser desorption and electrospray ionization (ESI). Originally developed for tissue imaging, it has significant potential for high throughput analysis of biological samples. Unlike MALDI, the matrix required for IR-MALDESI is water or ice, so no extra sample preparation is necessary for biological samples such as tissue, biofluids or buffered biochemical reactions. Decoupling the laser desorption process and allowing for post-desorption electrospray ionization limits ion suppression from salts and detergents, common additives for biochemical assay buffer systems, allowing biologically relevant buffers to be used with little concession to MS compatibility. Additionally, the theoretical limit on sampling rate is very high since laser desorption, electrospray ionization, and introduction of sample to the MS are achieved on the millisecond timescale, shifting the rate limiting steps to stage movement and MS data acquisition speed, which varies depending upon the mass analyzer type and operation mode. In addition to speed and compatibility with biological matrices IR-MALDESI can sample from any standard plate type and the cost of the ionization source (laser, stage, ESI source, etc.) is relatively inexpensive. This minimizes both consumable and capital expenses.

This talk will describe the implementation and optimization of IR-MALDESI for high throughput biochemical and cellular assays as well as high throughput analysis of protein post-translational modifications and protein-compound interactions. By coupling an IR-MALDESI ionization source to a Q Exactive HF-X mass spectrometer we have been able to achieve a 22 Hz sampling rate for biochemical and cellular assays while maintaining good quality assay Z’ of > 0.5 and %CV < 10%. In addition, whole proteins, up to 150 kDa, can be analyzed in < 1s and post-translational modifications or covalent compound additions monitored. Limited top-down sequencing of modification sites can also be performed on the Q Exactive instrument in short timeframes. Examples of each of these applications will be shown. In addition to the examples shown IR-MALDESI has potential to significantly increase productivity for ADME assays, PK/PD samples, permeability measurements, affinity-selection mass spectrometry, fermentation monitoring and multiple other applications in drug discovery. Its original develoment as an MS imaging technique also allows it to be used for imaging and solid surface analysis such as tablet degradation analysis, product QC, or counterfeit testing. 

Learning Objectives:

1. List three characteristics of an ideal high throughput assay detection technology.

2. Compare and contrast MALDESI-MS with LC-MS in terms of speed, versatility and sensitivity.

3. Discuss what experiments you could conduct with access to a MALDESI system.

Current drug discovery workflows rely on the synthesis and bioactivity assessment of large compound libraries, both processes which heavily depend on high-throughput (HT) experimentation to be able to generate and test hundreds of thousands of molecules as fast as possible. Due to the speed and versatility required for such testing, mass spectrometry (MS) has recently become a relevant alternative for HT analysis. However, despite MS being inherently fast, in many cases it still requires sample work-up (e.g. SPE), separation (e.g. LC) or extensive dilution, steps that might decrease throughput or sensitivity. 

This talk describes a novel HT system capable of (i) reaction screening, (ii) nano-scale synthesis, and (iii) label-free biological assays. This platform is based on desorption electrospray ionization (DESI), a contactless ambient ionization method (i.e. ionization occurs in the ambient environment with little or no sample preparation) with high tolerance to complex matrices. This DESI-MS system allows for analysis of high-density arrays (up to 6,144 samples per plate) with throughputs better than 1 sample per second and using only 50 nL samples (pg-ng amounts of material).  The platform is fully automated, capable of 24/7 continuous operation, and it automatically records MS and MS/MS spectra. The utility of this system to screen several organic reactions and identify optimal conditions for particular transformations or to study reaction mechanisms has been demonstrated. Similarly, HT DESI-MS has been proven useful to perform biological analyses, such as the monitoring of enzymatic activity directly from complex matrices (e.g. buffers with high detergent or salt concentrations), removing completely the need for sample work-up without any detriment to the quantitative performance (RSDs < 10%, LODs typically in the nM range). The concepts, workflow and applications of this new HT technology will be all overviewed during this presentation. 

Learning Objectives:

1. Describe the method of desorption electrospray ionization (DESI) and why is it useful for the rapid and direct analysis of complex samples.

2. Outline the general workflow of a high-throughput analysis using automated DESI-MS.

3. Illustrate some applications of high-throughput DESI-MS for reaction screening, nano-scale synthesis and biological assays.

This talk will focus on novel microfluidic approaches for measuring physical properties of particles in the 10 nm to 10 mm range and single cells, with particular focus on high precision measurement of mass, density, stiffness and the linking of these parameters to molecular readouts such as single-cell RNA sequencing. Ultimately, the ability to combine multi-parameter physical with molecular measurements at the single-cell level could not only be used to further understanding of important cellular processes but may also be used to increase the predictive power of clinical diagnostics. 

Learning Objectives:

1. Explain how single-cell measurements can be advantages for drug response assays.

2. Identify limitations of genomic-based biomarkers for personalized medicine in cancer.

3. Explain potential advantages of functional precision medicine for cancer patients.

When designing or implementing laboratory automation, we often have a fully automated end-goal in mind. We dream of leveraging automation to enable processes that run uninterrupted, preferably day and night. Achieving such fully automated workflows, however, is often a lengthy and challenging journey, where pitfalls along the way can result in frustration and a sense of unrealized potential. It is important to recognize though, that even when implemented only partially, or even minimally, automation already brings significant benefits to your organization.  Identifying and recognizing such intermediate gains is essential to achieve value realization early and throughout the automation journey. In this presentation I will showcase how automation (1) enables process standardization, (2) streamlines processes so that  bottlenecks can be avoided, (3) realizes assay miniaturization, (4) enables sophisticated design-of-experiments that are notoriously difficult to execute manually and (5) accounts for consistent data storage, along with its necessary metadata. I will demonstrate examples of these 5 benefits from Synthego's CRISPR genome engineering automation journey.  Finally, I will also highlight how parametrization and API interactions can help implement scalable automation solutions, specifically in light of recent focus on API support by major automation software and equipment vendors.

Learning Objectives:

1. Identify five values that lab automation brings, even when implemented only partially.

2. Discuss how automation enables experiments and processes that are not possible with a manual workflow.

3. Explain how APIs help to build scalable, parameterizable, lean workflows.